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1
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Velnar T, Kocivnik N, Bosnjak R. Clinical infections in neurosurgical oncology: An overview. World J Clin Cases 2023; 11:3418-3433. [PMID: 37383906 PMCID: PMC10294202 DOI: 10.12998/wjcc.v11.i15.3418] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 01/21/2023] [Revised: 03/05/2023] [Accepted: 04/13/2023] [Indexed: 05/25/2023] Open
Abstract
Central nervous system (CNS) infections are urgent conditions with high morbidity and mortality. Bacteria, viruses, parasites or fungi can cause them. Intracranial infections after craniotomies are an important complication of treatment, especially in oncological patients that are already immunologically compromised due to the disease and treatment. The consequence of CNS infections in oncological patients includes longer treatment with antibiotics, additional surgical procedures, higher treatment costs and poorer treatment outcomes. Additionally, the management of primary pathology may be prolonged or postponed as a result of the active infection. By introducing new and improved protocols, tightening controls on their implementation, constantly educating the entire team involved in patient treatment and educating both patients and relatives, the incidence of infections can be reduced effectively.
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Affiliation(s)
- Tomaz Velnar
- Department of Neurosurgery, University Medical Centre Ljubljana, Ljubljana 1000, Slovenia
- Alma Mater Europaea - ECM Maribor, Maribor 2000, Slovenia
| | - Nina Kocivnik
- Faculty of Pharmacy, University of Ljubljana, Ljubljana 1000, Slovenia
| | - Roman Bosnjak
- Department of Neurosurgery, University Medical Centre Ljubljana, Ljubljana 1000, Slovenia
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Xu J, Luo Y, Wang J, Tu W, Yi X, Xu X, Song Y, Tang Y, Hua X, Yu Y, Yin H, Yang Q, Huang WE. Artificial intelligence-aided rapid and accurate identification of clinical fungal infections by single-cell Raman spectroscopy. Front Microbiol 2023; 14:1125676. [PMID: 37032865 PMCID: PMC10073597 DOI: 10.3389/fmicb.2023.1125676] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/16/2022] [Accepted: 02/27/2023] [Indexed: 04/11/2023] Open
Abstract
Integrating artificial intelligence and new diagnostic platforms into routine clinical microbiology laboratory procedures has grown increasingly intriguing, holding promises of reducing turnaround time and cost and maximizing efficiency. At least one billion people are suffering from fungal infections, leading to over 1.6 million mortality every year. Despite the increasing demand for fungal diagnosis, current approaches suffer from manual bias, long cultivation time (from days to months), and low sensitivity (only 50% produce positive fungal cultures). Delayed and inaccurate treatments consequently lead to higher hospital costs, mobility and mortality rates. Here, we developed single-cell Raman spectroscopy and artificial intelligence to achieve rapid identification of infectious fungi. The classification between fungi and bacteria infections was initially achieved with 100% sensitivity and specificity using single-cell Raman spectra (SCRS). Then, we constructed a Raman dataset from clinical fungal isolates obtained from 94 patients, consisting of 115,129 SCRS. By training a classification model with an optimized clinical feedback loop, just 5 cells per patient (acquisition time 2 s per cell) made the most accurate classification. This protocol has achieved 100% accuracies for fungal identification at the species level. This protocol was transformed to assessing clinical samples of urinary tract infection, obtaining the correct diagnosis from raw sample-to-result within 1 h.
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Affiliation(s)
- Jiabao Xu
- Department of Engineering Science, University of Oxford, Oxford, United Kingdom
| | - Yanjun Luo
- Shanghai Hesen Biotech Co., Shanghai, China
| | - Jingkai Wang
- Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, China
| | - Weiming Tu
- Department of Engineering Science, University of Oxford, Oxford, United Kingdom
| | - Xiaofei Yi
- Institute of Antibiotics, Huashan Hospital, Fudan University, Shanghai, China
- National Clinical Research Center for Aging and Medicine, Huashan Hospital, Fudan University, Shanghai, China
| | - Xiaogang Xu
- Institute of Antibiotics, Huashan Hospital, Fudan University, Shanghai, China
- National Clinical Research Center for Aging and Medicine, Huashan Hospital, Fudan University, Shanghai, China
| | - Yizhi Song
- Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, China
| | - Yuguo Tang
- Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, China
| | - Xiaoting Hua
- Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, China
| | - Yunsong Yu
- Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, China
| | - Huabing Yin
- James Watt School of Engineering, University of Glasgow, Glasgow, United Kingdom
| | - Qiwen Yang
- Department of Clinical Laboratory, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
- *Correspondence: Qiwen Yang,
| | - Wei E. Huang
- Department of Engineering Science, University of Oxford, Oxford, United Kingdom
- Wei E. Huang,
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3
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Vandana UK, Barlaskar NH, Kalita R, Laskar IH, Mazumder PB. The Vital Foliar Diseases of Cicer arietinum L. (Chickpea): Science, Epidemiology, and Management. Fungal Biol 2020. [DOI: 10.1007/978-3-030-35947-8_10] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/29/2022]
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4
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Satyanarayana G. Work-up for Fever During Neutropenia for Both the Stem Cell Transplant Recipient and the Hematologic Malignancy Patient. Infect Dis Clin North Am 2019; 33:381-397. [PMID: 31005134 DOI: 10.1016/j.idc.2019.02.003] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/03/2023]
Abstract
Fever is a common complication in patients with underlying neutropenia and is associated with significant mortality in neutropenic patients with acute myelogenous leukemia or hematopoietic cell transplant. Fever may be the only sign of infection and requires further clinical assessment, including a history, a physical examination, and additional laboratory and radiographic testing. National and international guidelines recommend initiation of empiric antimicrobial therapy in patients with fever during neutropenia. Stepwise escalation of antibacterial therapy, followed by antifungal therapy for patients with persistent fever, generally is recommended. Consideration should also be given to de-escalation of antimicrobial therapy in the appropriate clinical settings.
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Affiliation(s)
- Gowri Satyanarayana
- Division of Infectious Diseases, Vanderbilt University Medical Center, A2200 MCN, 1161 21st Avenue South, Nashville, TN 37232-2605, USA.
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5
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Almeida-Silva F, Gonçalves DDS, de Abreu Almeida M, Guimarães AJ. Current Aspects of Diagnosis and Therapeutics of Histoplasmosis and Future Trends: Moving onto a New Immune (Diagnosis and Therapeutic) Era? CURRENT CLINICAL MICROBIOLOGY REPORTS 2019. [DOI: 10.1007/s40588-019-00118-3] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/18/2022]
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Hartl B, Zeller I, Manhart A, Selitsch B, Lass-Flörl C, Willinger B. A Retrospective Assessment of Four Antigen Assays for the Detection of Invasive Candidiasis Among High-Risk Hospitalized Patients. Mycopathologia 2018; 183:513-519. [PMID: 29356937 PMCID: PMC5958149 DOI: 10.1007/s11046-017-0238-1] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/29/2017] [Accepted: 12/15/2017] [Indexed: 11/04/2022]
Abstract
Because of their high mortality rates and non-specific symptoms, invasive Candida infections pose a huge diagnostic and therapeutic challenge. In this study, we evaluated the three mannan antigen assays Platelia, Platelia Plus and Serion, and the (1-3)-β-D-glucan assay Fungitell in a group of high-risk (hematological and surgical) patients. Test results of 305 patients hospitalized at the Vienna General Hospital and the University Hospital of Innsbruck were retrospectively analyzed. We assessed the test accuracy by means of descriptive statistics. Nine (2.95%) patients were affected by invasive candidiasis (IC), and 25 (8.2%) patients had a probable/possible infection. The majority of patients (271; 88.9%) showed no signs of infection. The Platelia and Serion mannan assays had a low sensitivity (65% and 52%, respectively), but high specificity (98% for both tests). The newer version of the Platelia assay, the Platelia Plus, had a higher sensitivity (85%) but a lower specificity (89%). The sensitivity of the Fungitell assay was high (100%), while its specificity was low (58%). The positive predictive values were 0.48 for the Platelia and 0.41 for the Serion assay, 0.26 for the Platelia Plus and 0.09 for the Fungitell assay. Our limited, retrospective study suggests the efficacy of mannan assays as screening (Platelia Plus) and confirmatory (Serion) tests, while the Fungitell assay can be used to exclude invasive Candida infections.
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Affiliation(s)
- Barbara Hartl
- Department of Laboratory Medicine, Division of Clinical Microbiology, Medical University of Vienna, Währinger Gürtel 18-20/5P, A-1090, Vienna, Austria
- Skånes universitetssjukhus, Getingevägen 4, 222 41, Lund, Sweden
| | - Iris Zeller
- Department of Laboratory Medicine, Division of Clinical Microbiology, Medical University of Vienna, Währinger Gürtel 18-20/5P, A-1090, Vienna, Austria
| | - Angelika Manhart
- Department of Laboratory Medicine, Division of Clinical Microbiology, Medical University of Vienna, Währinger Gürtel 18-20/5P, A-1090, Vienna, Austria
- Courant Institute of Mathematical Sciences, New York University, 251 Mercer Street, New York, NY, 10012, USA
| | - Brigitte Selitsch
- Department of Laboratory Medicine, Division of Clinical Microbiology, Medical University of Vienna, Währinger Gürtel 18-20/5P, A-1090, Vienna, Austria
| | - Cornelia Lass-Flörl
- Division of Hygiene and Medical Microbiology, Medical University of Innsbruck, Schöpfstrasse 41, A-6020, Innsbruck, Austria
| | - Birgit Willinger
- Department of Laboratory Medicine, Division of Clinical Microbiology, Medical University of Vienna, Währinger Gürtel 18-20/5P, A-1090, Vienna, Austria.
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Witkowska E, Jagielski T, Kamińska A. Genus- and species-level identification of dermatophyte fungi by surface-enhanced Raman spectroscopy. SPECTROCHIMICA ACTA. PART A, MOLECULAR AND BIOMOLECULAR SPECTROSCOPY 2018; 192:285-290. [PMID: 29156315 DOI: 10.1016/j.saa.2017.11.008] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 09/18/2017] [Revised: 10/16/2017] [Accepted: 11/02/2017] [Indexed: 06/07/2023]
Abstract
This paper demonstrates that surface-enhanced Raman spectroscopy (SERS) coupled with principal component analysis (PCA) can serve as a fast and reliable technique for detection and identification of dermatophyte fungi at both genus and species level. Dermatophyte infections are the most common mycotic diseases worldwide, affecting a quarter of the human population. Currently, there is no optimal method for detection and identification of fungal diseases, as each has certain limitations. Here, for the first time, we have achieved with a high accuracy, differentiation of dermatophytes representing three major genera, i.e. Trichophyton, Microsporum, and Epidermophyton. Two first principal components (PC), namely PC-1 and PC-2, gave together 97% of total variance. Additionally, species-level identification within the Trichophyton genus has been performed. PC-1 and PC-2, which are the most diagnostically significant, explain 98% of the variance in the data obtained from spectra of: Trichophyton rubrum, Trichophyton menatgrophytes, Trichophyton interdigitale and Trichophyton tonsurans. This study offers a new diagnostic approach for the identification of dermatophytes. Being fast, reliable and cost-effective, it has the potential to be incorporated in the clinical practice to improve diagnostics of medically important fungi.
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Affiliation(s)
- Evelin Witkowska
- Institute of Physical Chemistry, Polish Academy of Sciences, Kasprzaka 44/52, 01-224 Warsaw, Poland.
| | - Tomasz Jagielski
- University of Warsaw, Faculty of Biology, Institute of Microbiology, Department of Applied Microbiology, I. Miecznikowa 1, 02-096 Warsaw, Poland
| | - Agnieszka Kamińska
- Institute of Physical Chemistry, Polish Academy of Sciences, Kasprzaka 44/52, 01-224 Warsaw, Poland.
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Czurda S, Lion T. Broad-Spectrum Molecular Detection of Fungal Nucleic Acids by PCR-Based Amplification Techniques. Methods Mol Biol 2018; 1508:257-266. [PMID: 27837509 DOI: 10.1007/978-1-4939-6515-1_14] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 03/16/2023]
Abstract
Over the past decade, the incidence of life-threatening invasive fungal infections has dramatically increased. Infections caused by hitherto rare and emerging fungal pathogens are associated with significant morbidity and mortality among immunocompromised patients. These observations render the coverage of a broad range of clinically relevant fungal pathogens highly important. The so-called panfungal or, perhaps more correctly, broad-range nucleic acid amplification techniques do not only facilitate sensitive detection of all clinically relevant fungal species but are also rapid and can be applied to analyses of any patient specimens. They have therefore become valuable diagnostic tools for sensitive screening of patients at risk of invasive fungal infections. This chapter summarizes the currently available molecular technologies employed in testing of a wide range of fungal pathogens, and provides a detailed workflow for patient screening by broad-spectrum nucleic acid amplification techniques.
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Affiliation(s)
- Stefan Czurda
- Children's Cancer Research Institute (CCRI), St. Anna Kinderkrebsforschung, Vienna, Austria.,LabDia Labordiagnostik GmbH, Vienna, Austria
| | - Thomas Lion
- Children's Cancer Research Institute (CCRI), St. Anna Kinderkrebsforschung, Vienna, Austria. .,LabDia Labordiagnostik GmbH, Vienna, Austria. .,Department of Pediatrics, Medical University of Vienna, Vienna, Austria.
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9
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Rubinstein SM, Culos KA, Savani B, Satyanarayana G. Foiling fungal disease post hematopoietic cell transplant: review of prophylactic strategies. Bone Marrow Transplant 2017; 53:123-128. [PMID: 29058698 DOI: 10.1038/bmt.2017.222] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/13/2017] [Revised: 08/12/2017] [Accepted: 08/29/2017] [Indexed: 11/10/2022]
Abstract
Hematopoietic cell transplantation (HCT) offers definitive management for a wide variety of malignant and nonmalignant diseases. Conditioning regimens and therapies used to prevent and treat GvHD are immune suppressive, often increasing the risk of developing fungal disease due to yeasts or molds. Antifungal prophylaxis may be useful in preventing morbidity and mortality during and after HCT. In this article, we review the epidemiology and current literature regarding strategies for prevention of invasive fungal disease (IFD) in the pre-engraftment and post-engraftment settings, and propose future direction for scientific discovery.
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Affiliation(s)
- S M Rubinstein
- Department of Internal Medicine, Vanderbilt University Medical Center, Nashville, TN, USA
| | - K A Culos
- Department of Pharmaceutical Services, Vanderbilt University Medical Center, Nashville, TN, USA
| | - B Savani
- Division of Hematology/Oncology, Department of Internal Medicine, Vanderbilt University Medical Center, Nashville, TN, USA
| | - G Satyanarayana
- Division of Infectious Diseases, Department of Internal Medicine, Vanderbilt University Medical Center, Nashville, TN, USA
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10
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El-Aal AMA, El-Mashad N, Mohamed ASN. Revision on the Recent Diagnostic Strategies of Fungal Infections. OPEN JOURNAL OF MEDICAL MICROBIOLOGY 2017; 07:29-40. [DOI: 10.4236/ojmm.2017.71003] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 09/02/2023]
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Badiee P, Hashemizadeh Z, Ramzi M, Karimi M, Mohammadi R. Non-Invasive Methods to Diagnose Fungal Infections in Pediatric Patients with Hematologic Disorders. Jundishapur J Microbiol 2016; 9:e41573. [PMID: 28138379 PMCID: PMC5240159 DOI: 10.5812/jjm.41573] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/17/2016] [Revised: 10/08/2016] [Accepted: 10/09/2016] [Indexed: 12/15/2022] Open
Abstract
Background Invasive fungal infection (IFIs) is a major infectious complication in immunocompromised patients. Early diagnosis and initiation of antifungal therapy is important to achieve the best outcome. Objectives The current study aimed to investigate the incidence of IFIs and evaluate the diagnostic performance of non-invasive laboratory tests: serologic (β-D-glucan, galactomannan) and molecular (nested polymerase chain reaction) tests to diagnose fungal infections in hematologic pediatric patients. Patients and Methods In a cross-sectional study from October 2014 to January 2015, 321 blood samples of 62 pediatric patients with hematologic disorders and at high risk for fungal infections were analyzed. Non-invasive tests including the Platelia Aspergillus enzyme immunoassay (EIA) to detect galactomannan antigen, Glucatell for β–D–glucan and nested PCR to detect Candida and Aspergillus species-specific DNA were used in a weekly screening strategy. Results Twenty six patients (42%) were considered as proven and probable IFIs, including 3 (5%) proven and 23 (37%) probable cases. Eighteen patients (29%) were considered as possible cases. The sensitivity, specificity, positive and negative predictive values for galactomannan test in 26 patients with proven and probable fungal infections were 94.4%, 100%, 100% and 94.7%; for β-D-glucan test 92.3%, 77.7%, 85%, 87.5% and for nested-PCR were 84.6%, 88.8%, 91.7% and 80%, respectively. Conclusions The rate of IFIs in pediatric patients with hematologic disorders is high, and sample collection from the sterile sites cannot be performed in immunocompromised patients. Detection of circulating fungal cell wall components and DNA in the blood using non-invasive methods can offer diagnostic help in patients with suspected IFIs. Their results should be interpreted in combination with clinical, radiological and microbiological findings.
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Affiliation(s)
- Parisa Badiee
- Prof Alborzi Clinical Microbiology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Zahra Hashemizadeh
- Prof Alborzi Clinical Microbiology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
- Corresponding author: Zahra Hashemizadeh, PhD of Mycology, Prof Alborzi Clinical Microbiology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran. Tel: +98-36474303, Fax: +98-36474304, E-mail:
| | - Mani Ramzi
- Hematology Research Center, Transplant Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Mohammad Karimi
- Prof Alborzi Clinical Microbiology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Rasoul Mohammadi
- Department of Medical Parasitology and Mycology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
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12
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Thakur KT, Zunt JR. Approach to the international traveler with neurological symptoms. FUTURE NEUROLOGY 2015. [DOI: 10.2217/fnl.14.64] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/21/2022]
Abstract
ABSTRACT International travelers commonly contract illnesses while abroad, with the highest risk in those who spend extended time in developing countries. As travel to worldwide destinations becomes more accessible, neurologists should be aware of travel-related infections and noninfectious conditions presenting with neurological manifestations. Travelers may present with a myriad of neurologic symptoms, including confusion, headache, weakness and sensory symptoms. In this review, we discuss the general approach to the returning traveler with neurological symptoms and discuss the differential diagnosis of symptoms commonly encountered in practice.
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Affiliation(s)
- Kiran T Thakur
- Division of Neuroinfectious Disease & Neuroimmunology, Department of Neurology, Johns Hopkins Hospital, 600 North Wolfe Street, Meyer 6–113, Baltimore, MD 21205, USA
| | - Joseph R Zunt
- Department of Neurology, Global Health, Medicine (Infectious Diseases) & Epidemiology, University of Washington, Seattle, WA, USA
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13
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Molecular and nonmolecular diagnostic methods for invasive fungal infections. Clin Microbiol Rev 2015; 27:490-526. [PMID: 24982319 DOI: 10.1128/cmr.00091-13] [Citation(s) in RCA: 227] [Impact Index Per Article: 22.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/05/2023] Open
Abstract
Invasive fungal infections constitute a serious threat to an ever-growing population of immunocompromised individuals and other individuals at risk. Traditional diagnostic methods, such as histopathology and culture, which are still considered the gold standards, have low sensitivity, which underscores the need for the development of new means of detecting fungal infectious agents. Indeed, novel serologic and molecular techniques have been developed and are currently under clinical evaluation. Tests like the galactomannan antigen test for aspergillosis and the β-glucan test for invasive Candida spp. and molds, as well as other antigen and antibody tests, for Cryptococcus spp., Pneumocystis spp., and dimorphic fungi, have already been established as important diagnostic approaches and are implemented in routine clinical practice. On the other hand, PCR and other molecular approaches, such as matrix-assisted laser desorption ionization (MALDI) and fluorescence in situ hybridization (FISH), have proved promising in clinical trials but still need to undergo standardization before their clinical use can become widespread. The purpose of this review is to highlight the different diagnostic approaches that are currently utilized or under development for invasive fungal infections and to identify their performance characteristics and the challenges associated with their use.
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Khodadadi H, Mirhendi H, Makimura K, Satoh K, Karimi L, Izadi S. β-D-Glucan Assay in Diagnosis and Monitoring the Systemic Candidiasis in a Rat Model. Jundishapur J Microbiol 2014; 7:e10247. [PMID: 25371794 PMCID: PMC4217672 DOI: 10.5812/jjm.10247] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/13/2013] [Revised: 04/24/2013] [Accepted: 06/15/2013] [Indexed: 11/16/2022] Open
Abstract
BACKGROUND Determination of β-D-Glucan (BDG) in the serum aids to diagnose the invasive fungal infections. The current study evaluated the diagnostic potential value of BDG assay in monitoring the disease in experimental systemic candidiasis in a rat model. The results can provide a useful preliminary data to improve this approach in developing countries. OBJECTIVES The present study aimed to evaluate β-D-Glucan assay in diagnosis and monitoring the systemic candidiasis in a rat model. MATERIALS AND METHODS Twenty one rats were infected with 10(6) Candida albicans blastospore per rat. Twelve rats were considered as the negative controls (six immunocompromised rats without infection and six intact rats). During a week, every 24 hours the BDG sera level was determined by both Fungitell and Wako kits. To confirm the systemic infection in each rat, the suspensions of their internal organs were cultivated on agar plates and the number of colony forming units (CFU) of C. albicans was counted. RESULTS All the infected rats were positive with BDG tests. An increasing level of BDG was observed during early days after injection. The cutoff value for discrimination of BDG positive sera was obtained from the negative sera by the Fungitell kit. The sensitivity, specificity, positive and negative predictive values assessed for the Fungitell kit were 95%, 66.6%, 90.47% and 80%, respectively. These criteria for those of Wako were 90%, 83.3%, 94.7% and 71.4%, respectively. CONCLUSIONS While BDG assay seems to be a sensitive and specific adjunctive tool to diagnose and monitor the experimental systemic candidiasis, it seems that measuring the positive cutoff value in different laboratory conditions is necessary for favorable establishment of these tests.
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Affiliation(s)
- Hossein Khodadadi
- Department of Medical Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, IR Iran
| | - Hossein Mirhendi
- Department of Medical Parasitology and Mycology, School of Public Health, National Institute of Health Research, Tehran University of Medical Sciences, Tehran, IR Iran
- Corresponding author: Hossein Mirhendi, Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran. Tel/ Fax: +98-2188951392, E-mail:
| | - Koichi Makimura
- Institute of Medical Mycology, Teikyo University, Tokyo, Japan
| | - Kazuo Satoh
- Institute of Medical Mycology, Teikyo University, Tokyo, Japan
| | - Ladan Karimi
- Center of Medical Commission and Occupational Medicine, Social Security Organization, Esfahan, IR Iran
| | - Shahrokh Izadi
- Department of Medical Parasitology and Mycology, School of Public Health, National Institute of Health Research, Tehran University of Medical Sciences, Tehran, IR Iran
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15
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Romanelli AM, Fu J, Herrera ML, Wickes BL. A universal DNA extraction and PCR amplification method for fungal rDNA sequence-based identification. Mycoses 2014; 57:612-22. [PMID: 24865530 DOI: 10.1111/myc.12208] [Citation(s) in RCA: 36] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/19/2014] [Revised: 05/01/2014] [Accepted: 05/03/2014] [Indexed: 11/28/2022]
Abstract
Accurate identification of fungal pathogens using a sequence-based approach requires an extraction method that yields template DNA pure enough for polymerase chain reaction (PCR) or other types of amplification. Therefore, the objective of this study was to develop and standardise a rapid, inexpensive DNA extraction protocol applicable to the major fungal phyla, which would yield sufficient template DNA pure enough for PCR and sequencing. A total of 519 clinical and culture collection strains, comprised of both yeast and filamentous fungi, were prepared using our extraction method to determine its applicability for PCR, which targeted the ITS and D1/D2 regions in a single PCR amplicon. All templates were successfully amplified and found to yield the correct strain identification when sequenced. This protocol could be completed in approximately 30 min and utilised a combination of physical and chemical extraction methods but did not require organic solvents nor ethanol precipitation. The method reduces the number of tube manipulations and yielded suitable template DNA for PCR amplification from all phyla that were tested.
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Affiliation(s)
- A M Romanelli
- Department of Pathology and Laboratory Medicine, UC Davis Medical Center, Sacramento, CA, USA
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16
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Sánchez MJI, Pico AMP, Tejedor FM, Sánchez MJI, Acevedo RM. Using a polymerase chain reaction as a complementary test to improve the detection of dermatophyte fungus in nails. J Am Podiatr Med Assoc 2014; 104:233-7. [PMID: 24901581 DOI: 10.7547/0003-0538-104.3.233] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/03/2023]
Abstract
BACKGROUND Dermatomycoses are a group of pathologic abnormalities frequently seen in clinical practice, and their prevalence has increased in recent decades. Diagnostic confirmation of mycotic infection in nails is essential because there are several pathologic conditions with similar clinical manifestations. The classical method for confirming the presence of fungus in nail is microbiological culture and the identification of morphological structures by microscopy. METHODS We devised a nested polymerase chain reaction (PCR) that amplifies specific DNA sequences of dermatophyte fungus that is notably faster than the 3 to 4 weeks that the traditional procedure takes. We compared this new technique and the conventional plate culture method in 225 nail samples. The results were subjected to statistical analysis. RESULTS We found concordance in 78.2% of the samples analyzed by the two methods and increased sensitivity when simultaneously using the two methods to analyze clinical samples. Now we can confirm the presence of dermatophyte fungus in most of the positive samples in just 24 hours, and we have to wait for the result of culture only in negative PCR cases. CONCLUSIONS Although this PCR cannot, at present, substitute for the traditional culture method in the detection of dermatophyte infection of the nails, it can be used as a complementary technique because its main advantage lies in the significant reduction of time used for diagnosis, in addition to higher sensitivity.
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Affiliation(s)
| | - Ana María Pérez Pico
- Titulación de Podología, Centro Universitario de Plasencia, Universidad de Extremadura, Plasencia, Spain
| | - Félix Marcos Tejedor
- Titulación de Podología, Centro Universitario de Plasencia, Universidad de Extremadura, Plasencia, Spain
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Poissy J, Parmentier-Decrucq E, Sendid B, Mathieu D, Poulain D. Nouveaux marqueurs pour le diagnostic de la maladie fongique invasive. MEDECINE INTENSIVE REANIMATION 2014. [DOI: 10.1007/s13546-014-0866-4] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/29/2022]
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Cerqueira LB, de Francisco TMG, Gasparetto JC, Campos FR, Pontarolo R. Development and validation of an HPLC-MS/MS method for the early diagnosis of aspergillosis. PLoS One 2014; 9:e92851. [PMID: 24690884 PMCID: PMC3972208 DOI: 10.1371/journal.pone.0092851] [Citation(s) in RCA: 19] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/17/2013] [Accepted: 02/26/2014] [Indexed: 12/04/2022] Open
Abstract
Invasive aspergillosis is an opportunistic infection that is mainly caused by Aspergillus fumigatus, which is known to produce several secondary metabolites, including gliotoxin, the most abundant metabolite produced during hyphal growth. The diagnosis of invasive aspergillosis is often made late in the infection because of the lack of reliable and feasible diagnostic techniques; therefore, early detection is critical to begin treatment and avoid more serious complications. The present work reports the development and validation of an HPLC-MS/MS method for the detection of gliotoxin in the serum of patients with suspected aspergillosis. Chromatographic separation was achieved using an XBridge C18 column (150×2.1 mm id; 5 mm particle size) maintained at 25°C with the corresponding guard column (XBridge C18, 10×2.1 mm id, 5 mm particle size). The mobile phase was composed of a gradient of water and acetonitrile/water (95∶5 v/v), both containing 1 mM ammonium formate with a flow rate of 0.45 mL min−1. Data from the validation studies demonstrate that this new method is highly sensitive, selective, linear, precise, accurate and free from matrix interference. The developed method was successfully applied to samples from patients suspected of having aspergillosis. Therefore, the developed method has considerable potential as a diagnostic technique for aspergillosis.
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Affiliation(s)
| | | | - João C. Gasparetto
- Department of Pharmacy, Federal University of Paraná, Curitiba, Paraná, Brazil
| | | | - Roberto Pontarolo
- Department of Pharmacy, Federal University of Paraná, Curitiba, Paraná, Brazil
- * E-mail:
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Candida identification: a journey from conventional to molecular methods in medical mycology. World J Microbiol Biotechnol 2014; 30:1437-51. [PMID: 24379160 DOI: 10.1007/s11274-013-1574-z] [Citation(s) in RCA: 24] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/11/2013] [Accepted: 12/02/2013] [Indexed: 12/17/2022]
Abstract
The incidence of Candida infections have increased substantially in recent years due to aggressive use of immunosuppressants among patients. Use of broad-spectrum antibiotics and intravascular catheters in the intensive care unit have also attributed with high risks of candidiasis among immunocompromised patients. Among Candida species, C. albicans accounts for the majority of superficial and systemic infections, usually associated with high morbidity and mortality often caused due to increase in antimicrobial resistance and restricted number of antifungal drugs. Therefore, early detection of candidemia and correct identification of Candida species are indispensable pre-requisites for appropriate therapeutic intervention. Since blood culture based methods lack sensitivity, and species-specific identification by conventional method is time-consuming and often leads to misdiagnosis within closely related species, hence, molecular methods may provide alternative for accurate and rapid identification of Candida species. Although, several molecular approaches have been developed for accurate identification of Candida species but the internal transcribed spacer 1 and 2 (ITS1 and ITS2) regions of the rRNA gene are being used extensively in a variety of formats. Of note, ITS sequencing and PCR-RFLP analysis of ITS region seems to be promising as a rapid, easy, and cost-effective method for identification of Candida species. Here, we review a number of existing techniques ranging from conventional to molecular approaches currently in use for the identification of Candida species. Further, advantages and limitations of these methods are also discussed with respect to their discriminatory power, reproducibility, and ease of performance.
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Spanamberg A, Machado G, Casagrande RA, Sales GM, Fraga CF, Corbellini LG, Driemeier D, Ferreiro L. Aspergillus fumigatus from normal and condemned carcasses with airsacculitis in commercial poultry. PESQUISA VETERINÁRIA BRASILEIRA 2013. [DOI: 10.1590/s0100-736x2013000900004] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 11/22/2022]
Abstract
Carcass inspection is important for the detection of certain diseases and for monitoring their prevalence in slaughterhouses. The objective of this study was to assess the occurrence of aspergillosis caused by Aspergillus fumigatus in commercial poultry, through mycological and histopathological diagnosis, and to verify the causal association between the aspergillosis diagnosis criteria and condemnation due to airsacculitis in broilers through a case-control study. The study was carried out with 380 samples. Lungs were collected from broilers that were condemned (95) or not condemned (285) due to airsacculitis directly from the slaughter line. Forty-six (12%) lung samples were positive for A. fumigatus in mycological culture. Among all samples, 177 (46.6%) presented histopathological alterations, with necrotic, fibrinous, heterophilic pneumonia; heterophilic pneumonia and lymphoid hyperplasia being the most frequent. Out of the 380 lungs analyzed, 65.2% (30) showed histopathological alterations and isolation of fungi. The statistical analysis (McNemar's chi-square test) indicated a significant association between the presence of histopathological lesions and the isolation of A. fumigatus. Mycological cultivation and histopathological diagnosis increase the probability of detecting pulmonary alterations in birds condemned by the Final Inspection System, which suggests that such diagnostic criteria can improve the assessment and condemnation of birds affected by airsacculitis.
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Drago M, Scaltrito MM, Cariani L, Morace G. In VitroTesting ofAspergillus fumigatusClinical Isolates for Susceptibility to Voriconazole, Amphotericin B and Itraconazole: Comparison of Sensititre versus NCCLS M38-A Using Two Different Inocula. J Chemother 2013; 16:474-8. [PMID: 15565915 DOI: 10.1179/joc.2004.16.5.474] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/31/2022]
Abstract
Voriconazole, amphotericin B and itraconazole were tested in vitro against 18 strains of Aspergillus fumigatus isolated from cystic fibrosis patients. Susceptibility was tested with the broth microdilution method (M38-A protocol-NCCLS). Results of this reference method were compared with those of an experimental commercial microdilution broth method (Sensititre). Two different inocula, prepared from 2- and 7-day cultures, were used. Minimum inhibitory concentrations (MICs) of the reference method ranged from 0.25 to 2 microg/ml for voriconazole, 0.06 to 1 microg/ml for amphotericin B, 0.016 to >16 microg/ml for itraconazole. There were no significant differences in the MIC ranges or MIC90 values obtained with the two testing methods or with the two types of inocula. These findings confirm the good in vitro activity of voriconazole, itraconazole and amphotericin B against A. fumigatus. They also indicate that reliable susceptibility data can be generated more rapidly by commercial systems and use of 2-day cultures for inoculum preparation.
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Affiliation(s)
- M Drago
- Istituto di Microbiologia, Università degli Studi di Milano, Italy
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Jadhav S, Sahasrabudhe T, Kalley V, Gandham N. The microbial colonization profile of respiratory devices and the significance of the role of disinfection: a blinded study. JOURNAL OF CLINICAL AND DIAGNOSTIC RESEARCH : JCDR 2013; 7:1021-6. [PMID: 23905094 DOI: 10.7860/jcdr/2013/5681.3086] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [Subscribe] [Scholar Register] [Received: 01/23/2013] [Accepted: 03/24/2013] [Indexed: 11/24/2022]
Abstract
INTRODUCTION Approximately 10-40% of all the nosocomial infections are pulmonary, which lead to grave complications. Elderly, debilitated, or critically ill patients are at a high risk. The respiratory care equipments which include ventilators, humidifiers, nebulizers may have been identified as the potential vehicles which cause major nosocomial infections if they are colonized by fungi or bacteria. AIM To determine the rate of colonization by bacteria and fungi of the oxygen humidifier chambers of the portable cylinders and central lines at our hospital. The Hudson's chambers of nebulizers were also studied for the same. METHODS Swab samples were obtained from the equipments by using sterile cotton swabs on a tuesday, as these chambers were usually cleaned on every Saturday. Spot samples were taken from the ICUs, wards, the casualty and OPDs on a single day. Air samples were also obtained on the same day to determine whether the fungal spore load in the inhaled room air was normal or high. We performed a disinfection with 70% ethanol after cleaning these devices. RESULTS 53/70 (75.71%) samples showed fungal growth; out of which, 23/33 (69.70%) were from the ICU, 24/30(80%) were from the wards and 6/7 (85.71%) were from the OPDs. 23/30 (76.66%) swabs from the central line humidifiers, 18/23(78.26%) swabs from the O2 cylinder humidifiers and 8/17 (47.5%) swabs from the nebulizers grew bacteria. Of the total 61(87.14%) bacterial isolates, 42(68.85%) were gram negative bacteria and 19(31.14%) were gram positive cocci. Out of the 42 gram negative bacteria, 17 were multi-drug resistant like ESBL producers ie. Pseudomonas spp. (6) Acinetobacter spp.(4), Klebseilla pneumoniae (4), E.coli (2) and Stenotrophomonas maltophila (1). Our findings (before disinfection) showed that the colonization rate for fungi was 75% and that for bacteria, it was 87%. After the 70% ethanol disinfection and strict compliance with the hand hygiene, the colonization rates reduced significantly. The fungal colonization rate was reduced and only 15% fungi grew after the disinfection, while only 12% bacterial colonization rate was found. CONCLUSION This study indicates a potential in-hospital source of allergens and infections. The oxygen and nebulizer chambers need to be cleaned more frequently with disinfectants, to control the possible nosocomial infections.
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Affiliation(s)
- Savita Jadhav
- Associate Professor, Department of Microbiology, Pad. Dr. D.Y. Patil Medical College, Hospital & Research Centre (D.Y. Patil Vidyapeeth Pune) Pimpri-18. Maharashtra, India
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Morrissey CO. Advancing the Field: Evidence for New Management Strategies in Invasive Fungal Infections. CURRENT FUNGAL INFECTION REPORTS 2013; 7:51-58. [PMID: 23420637 PMCID: PMC3568482 DOI: 10.1007/s12281-012-0128-4] [Citation(s) in RCA: 12] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/07/2023]
Abstract
Invasive fungal infections (IFI) are a significant cause of morbidity and mortality in the immunocompromised. The traditional diagnostic methods of culture and histological examination lack sensitivity and often only make a diagnosis late when the fungal burden is high, reducing the chances of cure even with the availability of new more potent and less toxic antifungal agents. New non-culture-based serological and PCR assays have been developed. These appear more sensitive and are able to make an earlier diagnosis as compared with traditional diagnostic methods. Early diagnosis is central to reducing IFI-related morbidity and mortality. This review describes the diagnostic potential of the new serological and PCR assays and outlines how these assays have been incorporated into algorithms to improve the management of IFI.
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Affiliation(s)
- C. Orla Morrissey
- Infectious Diseases Unit, Alfred Health, Level 2, Burnet Building, 85 Commercial Road, Melbourne, Victoria 3004 Australia
- Department of Infectious Diseases, Central Clinical School, Monash University, Melbourne, Australia
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Abstract
The incidence of invasive fungal infections (IFIs) has seen a marked increase in the last two decades. This is especially evident among transplant recipients, patients suffering from AIDS, in addition to those in receipt of immunosuppressive therapy. Worryingly, this increased incidence includes infections caused by opportunistic fungi and emerging fungal infections which are resistant to or certainly less susceptible than others to standard antifungal agents. As a direct response to this phenomenon, there has been a resolute effort over the past several decades to improve early and accurate diagnosis and provide reliable screening protocols thereby promoting the administration of appropriate antifungal therapy for fungal infections. Early diagnosis and treatment with antifungal therapy are vital if a patient is to survive an IFI. Substantial advancements have been made with regard to both the diagnosis and subsequent treatment of an IFI. In parallel, stark changes in the epidemiological profile of these IFIs have similarly occurred, often in direct response the type of antifungal agent being administered. The effects of an IFI can be far reaching, ranging from increased morbidity and mortality to increased length hospital stays and economic burden.
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Affiliation(s)
- Nina L Tuite
- Molecular Diagnostics Research Group, National University of Ireland, Galway, Ireland.
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Detection, identification, and distribution of fungi in bronchoalveolar lavage specimens by use of multilocus PCR coupled with electrospray ionization/mass spectrometry. J Clin Microbiol 2012; 51:136-41. [PMID: 23100337 DOI: 10.1128/jcm.01907-12] [Citation(s) in RCA: 44] [Impact Index Per Article: 3.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/15/2023] Open
Abstract
As pulmonary fungal infections continue to increase due to an increasing number of immunocompromised patients, rapid detection and accurate identification of these fungal pathogens are critical. A broad fungal assay was developed by incorporating broad-range multilocus PCR amplification and electrospray ionization/mass spectrometry (PCR/ESI-MS) to detect and identify fungal organisms directly from clinical specimens. The aims of this study were to evaluate the performance of PCR/ESI-MS for detection, identification, and determination of the distribution of fungal organisms in bronchoalveolar lavage (BAL) fluid specimens. The BAL fluid specimens submitted for fungal culture at Vanderbilt University Medical Center between May 2005 and October 2011 were included. Cultures and identification were done using standard procedures. In addition, DNA was extracted from BAL fluid specimens, and fungal DNA amplification/identification were performed by PCR/ESI-MS. The results were compared with those of the standard cultures. A total of 691 nonduplicated BAL fluid specimens with sufficient leftover volume for molecular testing were evaluated using PCR/ESI-MS. Among them, 134 specimens (19.4%) were positive for fungi by both culture and PCR/ESI-MS testing. Of the dual-positive specimens, 125 (93.3%) were positive for Candida and Aspergillus species, with concordances between culture and PCR/ESI-MS results being 84 (67.2%) at the species level and 109 (87.2%) at the genus level. In addition, 243 (35.2%) and 30 (4.3%) specimens were positive only by PCR/ESI-MS or by culture, respectively (odds ratio [OR] = 11.95, 95% confidence interval [CI] = 7.90 to 18.17, P = 0.0000). Codetection of fungal organisms was noted in 23 (3.3%) specimens by PCR/ESI-MS, which was significantly higher than the 4 (0.6%) in which they were noted by culture (OR = 5.91, 95% CI = 1.93 to 20.27, P = 0.0002). Among 53 specimens in which cultures failed because of bacterial overgrowth, at least one fungus was identified in 26 specimens (47.3%) by PCR/ESI-MS. PCR/ESI-MS provides an advanced tool for rapid and sensitive detection, identification, and determination of the distribution of fungal organisms directly from BAL fluid specimens. Moreover, it detected fungal organisms in specimens in which cultures failed because of bacterial overgrowth. The clinical relevance of the significantly higher detection rate of fungal organisms by PCR/ESI-MS merits further investigation.
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Arunmozhi Balajee S, Hurst SF, Chang LS, Miles M, Beeler E, Hale C, Kasuga T, Benedict K, Chiller T, Lindsley MD. Multilocus sequence typing of Histoplasma capsulatum in formalin-fixed paraffin-embedded tissues from cats living in non-endemic regions reveals a new phylogenetic clade. Med Mycol 2012; 51:345-51. [PMID: 23072593 DOI: 10.3109/13693786.2012.733430] [Citation(s) in RCA: 23] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022] Open
Abstract
Infections caused by Histoplasma capsulatum are found most often in endemic regions of North, Central, and South America. H. capsulatum has been divided into eight geographic clades by multi-locus sequence typing (MLST). Recently, one isolate and five formalin-fixed paraffin-embedded (FFPE) tissue samples were received from six of 15 suspected cases of histoplasmosis in cats residing in areas not known to be endemic for H. capsulatum. Polymerase chain reaction (PCR) amplification and sequence analysis of the rDNA ITS-2 region confirmed the diagnosis of H. capsulatum. Since these cases were not, as noted, from the accepted endemic areas, it was of interest to understand the molecular epidemiology of these isolates. Results of molecular analysis indicated that the H. capsulatum recovered from the cats were most closely related to the North American-1 clade, but clustered separately outside this clade, suggesting that the H. capsulatum infecting the animals may represent a separate clade or phylogenetic species. This study also demonstrated the utility of obtaining valuable molecular subtype data directly from archived FFPE tissue blocks, particularly when a fungus culture was not performed or is otherwise unavailable.
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Affiliation(s)
- S Arunmozhi Balajee
- Center For Global Health , Centers for Disease Control and Prevention, Atlanta, GA 30333, USA
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Kadmon G, Nahum E, Sprecher H, Stein J, Levy I, Schiller O, Schonfeld T. Polymerase-chain-reaction-based diagnosis of invasive fungal pulmonary infections in immunocompromised children. Pediatr Pulmonol 2012; 47:994-1000. [PMID: 22328487 DOI: 10.1002/ppul.22523] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 10/21/2011] [Accepted: 12/04/2011] [Indexed: 11/06/2022]
Abstract
OBJECTIVE Fungal pneumonia is a serious complication in immunocompromised children. It is difficult to diagnose because of the low sensitivity of clinical and standard laboratory tests. The aim of this study was to investigate the diagnostic impact of polymerase chain reaction (PCR) assays for fungal pathogens in bronchoalveolar lavage (BAL) fluid. STUDY DESIGN BAL samples obtained from hospitalized immunocompromised patients with clinical pneumonia between January 2007 and June 2009 were processed for microscopy and cultures in addition to PCR-based fungal assays. The results were compared between the standard and PCR methods. RESULTS Seventy-seven children with 100 episodes of pneumonia were included in the study. Fungal pathogens were detected by standard microbiological investigations in 10 episodes (10%) and by PCR-based assays alone in 20 episodes (20%). There was no significant difference in clinical improvement or mortality rate between patients diagnosed by the different methods. In 61 episodes, no fungal pathogen was identified by either method. Prolonged antifungal therapy was avoided in 43 episodes. CONCLUSION PCR-based assay for the diagnosis of fungal pulmonary infections may be a useful adjunct to clinical and standard microbiological techniques. The use of PCR may decrease the time to diagnosis, increase the rate of detection of fungal pathogens, and spare patients unnecessary antifungal treatment.
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Affiliation(s)
- Gili Kadmon
- Pediatric Intensive Care Unit, Schneider Children's Medical Center of Israel, Petach Tikva, Israel.
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Buelow DR, Gu Z, Walsh TJ, Hayden RT. Evaluation of multiplexed PCR and liquid-phase array for identification of respiratory fungal pathogens. Med Mycol 2012; 50:775-80. [PMID: 22435876 DOI: 10.3109/13693786.2012.666681] [Citation(s) in RCA: 18] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022] Open
Abstract
Invasive fungal infections are the cause of serious morbidity and high mortality in immunocompromised patients. Early laboratory diagnostic options remain limited; however, rapid detection and accurate identification may improve outcome. Herein, multiplexed PCR followed by liquid-phase array was evaluated for detection and identification of common respiratory fungal pathogens, including Aspergillus fumigatus, Rhizopus microsporus, Scedosporium apiospermum and Fusarium solani. The limit of detection ranged 0.1-1 ng of DNA, depending on the fungus being tested. Primer cross-reactivity was seen for some fungi: Aspergillus flavus primers detected Aspergillus oryzae; Scedosporium apiospermum primers detected Paecilomyces lilacinus, and Aspergillus terreus primers detected S. apiospermum. PCR followed by liquid-phase array is potentially useful for the identification of clinically relevant fungal pathogens.
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Affiliation(s)
- Daelynn R Buelow
- Department of Pathology, St Jude Children's Research Hospital, Memphis, TN 38105 - 2794, USA
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Development of specific sequence-characterized amplified region markers for detecting Histoplasma capsulatum in clinical and environmental samples. J Clin Microbiol 2011; 50:673-9. [PMID: 22189121 DOI: 10.1128/jcm.05271-11] [Citation(s) in RCA: 18] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022] Open
Abstract
Sequence-characterized amplified region (SCAR) markers, generated by randomly amplified polymorphic DNA (RAPD)-PCR, were developed to detect Histoplasma capsulatum selectively in clinical and environmental samples. A 1,200-bp RAPD-PCR-specific band produced with the 1281-1283 primers was cloned, sequenced, and used to design two SCAR markers, 1281-1283(220) and 1281-1283(230). The specificity of these markers was confirmed by Southern hybridization. To evaluate the relevance of the SCAR markers for the diagnosis of histoplasmosis, another molecular marker (M antigen probe) was used for comparison. To validate 1281-1283(220) and 1281-1283(230) as new tools for the identification of H. capsulatum, the specificity and sensitivity of these markers were assessed for the detection of the pathogen in 36 clinical (17 humans, as well as 9 experimentally and 10 naturally infected nonhuman mammals) and 20 environmental (10 contaminated soil and 10 guano) samples. Although the two SCAR markers and the M antigen probe identified H. capsulatum isolates from different geographic origins in America, the 1281-1283(220) SCAR marker was the most specific and detected the pathogen in all samples tested. In contrast, the 1281-1283(230) SCAR marker and the M antigen probe also amplified DNA from Aspergillus niger and Cryptococcus neoformans, respectively. Both SCAR markers detected as little as 0.001 ng of H. capsulatum DNA, while the M antigen probe detected 0.5 ng of fungal DNA. The SCAR markers revealed the fungal presence better than the M antigen probe in contaminated soil and guano samples. Based on our results, the 1281-1283(220) marker can be used to detect and identify H. capsulatum in samples from different sources.
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Abstract
Sporotrichosis, which is caused by the dimorphic fungus Sporothrix schenckii, is currently distributed throughout the world, especially in tropical and subtropical zones. Infection generally occurs by traumatic inoculation of soil, plants, and organic matter contaminated with the fungus. Certain leisure and occupational activities, such as floriculture, agriculture, mining, and wood exploitation, are traditionally associated with the mycosis. Zoonotic transmission has been described in isolated cases or in small outbreaks. Since the end of the 1990s there has been an epidemic of sporotrichosis associated with transmission by cats in Rio de Janeiro, Brazil. More than 2,000 human cases and 3,000 animal cases have been reported. In humans, the lesions are usually restricted to the skin, subcutaneous cellular tissue, and adjacent lymphatic vessels. In cats, the disease can evolve with severe clinical manifestations and frequent systemic involvement. The gold standard for sporotrichosis diagnosis is culture. However, serological, histopathological, and molecular approaches have been recently adopted as auxiliary tools for the diagnosis of this mycotic infection. The first-choice treatment for both humans and cats is itraconazole.
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Hsu JL, Ruoss SJ, Bower ND, Lin M, Holodniy M, Stevens DA. Diagnosing invasive fungal disease in critically ill patients. Crit Rev Microbiol 2011; 37:277-312. [PMID: 21749278 DOI: 10.3109/1040841x.2011.581223] [Citation(s) in RCA: 49] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/31/2022]
Abstract
Fungal infections are increasing, with a changing landscape of pathogens and emergence of new groups at risk for invasive disease. We review current diagnostic techniques, focusing on studies in critically ill patients. Microbiological cultures, the current "gold standard", demonstrate poor sensitivity, thus diagnosis of invasive disease in the critically ill is difficult. This diagnostic dilemma results in under- or over-treatment of patients, potentially contributing to poor outcomes and antifungal resistance. While other current diagnostic tests perform moderately well, many lack timeliness, efficacy, and are negatively affected by treatments common to critically ill patients. New nucleic acid-based research is promising.
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Affiliation(s)
- Joe L Hsu
- Division of Pulmonary and Critical Care Medicine, Stanford University School of Medicine, Stanford, CA, USA
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PARK DH, PARK K, PARK J, PARK HH, CHAE H, LIM J, OH EJ, KIM Y, PARK YJ, HAN K. Screening of sepsis using leukocyte cell population data from the Coulter automatic blood cell analyzer DxH800. Int J Lab Hematol 2011; 33:391-9. [DOI: 10.1111/j.1751-553x.2011.01298.x] [Citation(s) in RCA: 47] [Impact Index Per Article: 3.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/26/2022]
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Chen S, Erhart LM, Anderson S, Komatsu K, Park B, Chiller T, Sunenshine R. Coccidioidomycosis: knowledge, attitudes, and practices among healthcare providers--Arizona, 2007. Med Mycol 2011; 49:649-56. [PMID: 21247229 DOI: 10.3109/13693786.2010.547995] [Citation(s) in RCA: 21] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022] Open
Abstract
Coccidioidomycosis presumably causes ≤ 33% of community-acquired pneumonias cases, although < 15% of the patients are tested for coccidioidomycosis. We assessed healthcare providers' knowledge, attitudes, and practices regarding coccidioidomycosis diagnosis and treatment in Arizona. A survey was mailed to 7,608 eligible healthcare providers licensed by the Arizona medical, osteopathic, and nursing boards in October and December 2007. We used weights to adjust for non-response and multivariate logistic regression models to identify predictors of ≥ 70% correct regarding knowledge and treatment practices. Of 1,823 (24%) respondents, 53% were physicians, 52% were male, and the mean age was 51 years. Approximately 50% reported confidence in their ability to treat coccidioidomycosis, and 21% correctly answered all four treatment questions. Predictors of ≥ 70% correct concerning knowledge and treatment practices included always counseling patients after diagnosis (adjusted odds ratio [AOR]=4.4; 95% confidence interval [CI]: 2.8-7.1); specializing in infectious diseases (AOR=2.4; 95% CI: 1.0-5.7); and having received coccidioidomycosis continuing medical education (CME) in the last year (AOR=1.8; 95% CI: 1.2-2.6). These findings demonstrate that coccidioidomycosis CME improves knowledge of disease diagnosis and management, underscoring the need for a comprehensive coccidioidomycosis education campaign for healthcare providers in Arizona.
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Affiliation(s)
- Sanny Chen
- Epidemic Intelligence Service, Centers for Disease Control and Prevention (CDC), Atlanta, Georgia 30333, USA.
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Chandrasekar P. Diagnostic challenges and recent advances in the early management of invasive fungal infections. Eur J Haematol 2010; 84:281-90. [DOI: 10.1111/j.1600-0609.2009.01391.x] [Citation(s) in RCA: 35] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/15/2023]
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Tanriover MD, Ascioglu S, Altun B, Uzun O. Galactomannan on the stage: prospective evaluation of the applicability in routine practice and surveillance. Mycoses 2010; 53:16-25. [PMID: 20091935 DOI: 10.1111/j.1439-0507.2008.01652.x] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/29/2022]
Abstract
Invasive aspergillosis (IA) presents a diagnostic and therapeutic dilemma for the physicians who take care of the patients with severe underlying diseases and immunosuppression. This study aimed to evaluate the usefulness of serum galactomannan (GM) measurements in the routine practice and surveillance of IA along with possible caveats in diagnosis and treatment. Adult patients with high-risk haematological malignancies admitted to the Internal Medicine wards during the 2-year study period were followed up by daily visits for vital signs, existing or newly developing signs and symptoms, clinical and laboratory findings. Blood samples were analysed for GM levels by the ELISA method at the end of the study period. Data of 58 hospitalisation episodes in 45 patients were analysed. Proven IA was diagnosed in one patient, probable IA was diagnosed in four patients. The sensitivity was 60% and the specificity was 21% when the index cut-off for positivity was accepted as 0.5. The yield of GM testing may be influenced by many variables and each centre should evaluate the usefulness of this test in its own conditions.
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Affiliation(s)
- Mine Durusu Tanriover
- Department of Internal Medicine, Faculty of Medicine, Hacettepe University, Ankara, Turkey.
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Abstract
PURPOSE OF REVIEW Antifungal drug resistance is a confounding factor that negatively impacts clinical outcome for patients with serious mycoses. Early detection of fungi in blood or other specimens with a rapid assessment of drug susceptibility could improve the survival of patients with invasive disease by accelerating the initiation of appropriate antifungal treatment. Recent years have seen the growth of molecular technology that is ideally suited for fungal identification and assessment of drug resistance mechanisms. RECENT FINDINGS Elucidation of the genetic mechanisms responsible for triazole and echinocandin resistance in prominent Candida spp. and Aspergillus spp. provides an opportunity to develop molecular diagnostic platforms suitable for rapid detection of primary and secondary drug resistance. Several highly dynamic and robust amplification/detection methodologies are now available that can provide simultaneous species identification and high fidelity discrimination of resistance alleles. SUMMARY Molecular diagnostic platforms are ideal for rapid detection of fungal pathogens and they provide an opportunity to develop in parallel molecular assays that can evaluate antifungal drug resistance.
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Sequence-based identification of filamentous basidiomycetous fungi from clinical specimens: a cautionary note. J Clin Microbiol 2009; 48:741-52. [PMID: 20042628 DOI: 10.1128/jcm.01948-09] [Citation(s) in RCA: 94] [Impact Index Per Article: 5.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022] Open
Abstract
The species-level identification of sterile and/or arthroconidium-forming filamentous fungi presumed to be basidiomycetes based upon morphological or physiological features alone is usually not possible due to the limited amount of hyphal differentiation. Therefore, a reliable molecular approach capable of the unambiguous identification of clinical isolates is needed. One hundred sixty-eight presumptive basidiomycetes were screened by sequence analysis of the internal transcribed spacer (ITS) and D1/D2 ribosomal DNA regions in an effort to obtain a species identification. Through the use of this approach, identification of a basidiomycetous fungus to the species level was obtained for 167/168 of the isolates. However, comparison of the BLAST results for each isolate for both regions revealed that only 28.6% (48/168) of the isolates had the same species identification by use of both the ITS and the D1/D2 regions, regardless of the percent identity. At the less stringent genus-only level, the identities for only 48.8% (82/168) of the isolates agreed for both regions. Investigation of the causes for this low level of agreement revealed that 14% of the species lacked an ITS region deposit and 16% lacked a D1/D2 region deposit. Few GenBank deposits were found to be complete for either region, with only 8% of the isolates having a complete ITS region and 10% having a complete D1/D2 region. This study demonstrates that while sequence-based identification is a powerful tool for many fungi, sequence data derived from filamentous basidiomycetes should be interpreted carefully, particularly in the context of missing or incomplete GenBank data, and, whenever possible, should be evaluated in light of compatible morphological features.
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Xavier MO, Oliveira FDM, Severo LC. Capítulo 1: diagnóstico laboratorial das micoses pulmonares. J Bras Pneumol 2009; 35:907-19. [DOI: 10.1590/s1806-37132009000900013] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022] Open
Abstract
Nesta era de imunossupressão e transplantes, é imperativa a comunicação entre médicos e laboratoristas devido ao fato de que o diagnóstico de doenças fúngicas, para esses pacientes, deve ser rápido, o que é complicado e requer a cooperação e colaboração de vários profissionais com distintas especializações. Este artigo revisa as técnicas laboratoriais utilizadas para o diagnóstico de infecções fúngicas pulmonares. Os tópicos abordados incluem: fatores relacionados ao hospedeiro, como resposta imunológica e predisposições anatômicas; colheita, armazenamento, remessa e transporte das amostras; processamento laboratorial; exame microscópico direto; técnicas de coloração, cultivo e identificação fúngica; biossegurança em laboratórios; tropismo e reação teciduais; soromicologia; e detecção de antígenos.
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Raymond S, Henon T, Grenouillet F, Legrand F, Woronoff-Lemsi MC, Hoen B, Limat S, Leroy J. Audit clinique des prescriptions d’antifongiques systémiques couteux au centre hospitalier universitaire de Besançon. Med Mal Infect 2009; 39:125-32. [DOI: 10.1016/j.medmal.2008.09.028] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/09/2008] [Revised: 07/23/2008] [Accepted: 09/30/2008] [Indexed: 10/21/2022]
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Apaire-Marchais V, Kempf M, Lefrançois C, Marot A, Licznar P, Cottin J, Poulain D, Robert R. Evaluation of an immunomagnetic separation method to capture Candida yeasts cells in blood. BMC Microbiol 2008; 8:157. [PMID: 18808691 PMCID: PMC2556679 DOI: 10.1186/1471-2180-8-157] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/19/2008] [Accepted: 09/22/2008] [Indexed: 11/10/2022] Open
Abstract
BACKGROUND Candida species have become the fourth most-frequent cause of nosocomial bloodstream infections in immunocompromised patients. Therefore, rapid identification of pathogenic fungi to species level has been considered critical for treatment. Conventional diagnostic procedures such as blood culture or biochemical tests are lacking both sensitivity and species specificity, so development of rapid diagnostic is essential. RESULTS An immunomagnetic method involving anti-Candida monoclonal antibodies was developed to capture and concentrate in human blood four different species of Candida cells responsible for invasive yeast infections. In comparison with an automated blood culture, processing time of immunomagnetic separation is shorter, saving at least 24 hours to obtain colonies before identification. CONCLUSION Thus, this easy to use method provides a promising basis for concentrating all Candida species in blood to improve sensitivity before identification.
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Affiliation(s)
- Véronique Apaire-Marchais
- Groupe d'Etude des Interactions Hôte-Parasite, UPRES EA 3142, UFR des Sciences Pharmaceutiques et d'Ingénierie de la Santé, Angers Cedex, France.
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Mirkov I, Zolotarevski L, Glamočlija J, Kataranovski D, Kataranovski M. Experimental disseminated aspergillosis in mice: Histopathological study. J Mycol Med 2008. [DOI: 10.1016/j.mycmed.2008.02.001] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/22/2022]
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Cruciani M, Serpelloni G. Management of Candida infections in the adult intensive care unit. Expert Opin Pharmacother 2008; 9:175-91. [PMID: 18201143 DOI: 10.1517/14656566.9.2.175] [Citation(s) in RCA: 18] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/05/2022]
Abstract
The epidemiology of Candida infection in intensive care units (ICUs) and the management strategies for such infections in non-neutropenic intensive care patients are discussed in this review. Candida species are one of the leading causes of nosocomial bloodstream infections and a significant cause of morbidity in patients admitted to the ICU. Prophylactic, pre-emptive and empiric treatment strategies for Candida infections have been explored in ICU patients. Routine prophylaxis should not be administered to the whole population of ICU patients, because the concerns about the selection of azole-resistant Candida strains or the induction of resistance are justified. Treatment of fungal infections is now possible with newer antifungal agents, including newer azoles (e.g., voriconazole, posaconazole) and echinocandins (e.g., micafungin, anidulafungin). However, there is a critical need for improvement in diagnosis of invasive Candida infection in order to provide clinicians the opportunity to intervene earlier in the diseases course.
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Affiliation(s)
- Mario Cruciani
- Center of Preventive Medicine & HIV Out-Patient Clinic, V. Germania, 20-37135 Verona, Italy.
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45
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46
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Abstract
This chapter discusses the anatomy, functions, and biochemistry of cerebrospinal fluid (CSF). CSF has four major functions: physical support of neural structures, excretion and “sink” action, intracerebral transport, and control of the chemical environment of the central nervous system. CSF provides a “water jacket” of physical support and buoyancy. The CSF is protective because its volume changes reciprocally with changes in the volume of intracranial contents, particularly blood. Thus, the CSF protects the brain from changes in arterial and central venous pressure associated with posture, respiration, and exertion. Acute or chronic pathological changes in intracranial contents can be accommodated, to a point, by changes in the CSF volume. The direct transfer of brain metabolites into the CSF provides excretory function. This capacity is important because the brain lacks a lymphatic system. The lymphatic function of the CSF is also manifested in the removal of large proteins and cells, such as bacteria or blood cells, by bulk CSF absorption. The “sink” action of the CSF arises from the restricted access of water-soluble substances to the CSF and the low concentration of these solutes in the CSF.
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Kedzierska A, Kochan P, Pietrzyk A, Kedzierska J. Current status of fungal cell wall components in the immunodiagnostics of invasive fungal infections in humans: galactomannan, mannan and (1-->3)-beta-D-glucan antigens. Eur J Clin Microbiol Infect Dis 2007; 26:755-66. [PMID: 17671803 DOI: 10.1007/s10096-007-0373-6] [Citation(s) in RCA: 59] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/16/2022]
Abstract
Early diagnosis of fungal infections and the implementation of appropriate treatment represent major issues for clinicians, nowadays. Histopathological demonstration of microorganisms in tissue specimens or growth of fungal agents in culture media is still considered the "gold standard", but obtaining such specimens may be difficult. Several groups have investigated serological assays for cell wall elements unique to fungal organisms in serum or other body fluids to improve diagnostics in patients with haematological malignancies or undergoing haematopoietic stem-cell transplantation. In this review we have concentrated on the currently available assays allowing for detection of highly immunogenic components of fungal cell wall: galactomannan, mannan, and also (1-->3)-beta-D-glucan. Rapid serological tests appear to be useful for screening high-risk haematological patients, since they allow for the early diagnosis of invasive fungal infections, including infections with the most common pathogens such as Aspergillus and Candida. Based on current literature, factors increasing the probability of obtaining false-positive or false-negative results detected by each test were also analysed and tabulated.
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Affiliation(s)
- A Kedzierska
- Department of Clinical Microbiology, Chair of Clinical Immunology and Transplantology, Polish-American Institute of Paediatrics, Jagiellonian University Medical College, 265 Wielicka Street, 30-663, Cracow, Poland
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Yoo JH, Choi SM, Lee DG, Park SH, Choi JH, Kwon EY, Shin WS. Comparison of the real-time nucleic acid sequence-based amplification (RTi-NASBA) with conventional NASBA, and galactomannan assay for the diagnosis of invasive aspergillosis. J Korean Med Sci 2007; 22:672-6. [PMID: 17728508 PMCID: PMC2693818 DOI: 10.3346/jkms.2007.22.4.672] [Citation(s) in RCA: 22] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 11/25/2022] Open
Abstract
We compared a real time-nucleic acid sequence-based amplification (RTi-NASBA) with conventional NASBA, galactomannan enzyme immunosorbent assay (GMEIA), and Mycology Study Group of the European Organization for Research and Treatment of Cancer (EORTC/MSG) criteria for the diagnosis of invasive aspergillosis (IA). From May 2004 to May 2005, blood samples (314 in total) were collected twice a week from 78 patients with hematologic diseases during neutropenic fever after chemotherapy or hematopoietic stem cell transplantation. Results were compared with each other on the basis of EORTC/ MSG criteria. The cutoff of conventional NASBA was set to be 3.5; GM 0.5; RTi-NASBA, 20% above the negative control. There were 22 patients with IA (7 probables and 15 possibles) and 56 patients with nonfungal infection. The Kappa statistic for RTi-NASBA versus conventional NASBA was 0.80 (0.66-0.82; p<0.001) indicating that there was fairly good accordance between two tests. RTi-NASBA showed sensitivity 0.96, specificity 0.43, positive- and negative-predictive value 0.40 and 0.96, respectively. GM showed good specificity (0.98), while the sensitivity (0.45) was poor. When we use the combination of GM with either of two NASBAs, the sensitivity was improved up to 100%. In conclusion, RTi-NASBA could be a good alternative to the conventional one for the screening of IA.
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Affiliation(s)
- Jin Hong Yoo
- Department of Internal Medicine, Holy Family Hospital, Sosa-2-dong, Wonmi-gu, Bucheon 420-717, Korea.
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Maertens J, Theunissen K, Lodewyck T, Lagrou K, Van Eldere J. Advances in the serological diagnosis of invasive Aspergillus infections in patients with haematological disorders. Mycoses 2007; 50 Suppl 1:2-17. [PMID: 17394605 DOI: 10.1111/j.1439-0507.2007.01375.x] [Citation(s) in RCA: 58] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/30/2022]
Abstract
A reliable diagnosis of invasive aspergillosis in patients with haematological malignancies is seldom achieved antemortem. Conventional laboratory diagnostic methods are insensitive and time-consuming, resulting in late diagnosis and treatment and contributing to unacceptably high mortality. As a result, routine antifungal prophylaxis and early empirical treatment have been recommended. However, overtreatment associated with these strategies results in increased toxicity and cost. The use of sensitive and rapid non-culture-based diagnostic assays, such as detection of Aspergillus antigens (galactomannan, beta-D-glucan) or detection of genomic DNA sequences may allow a shift in emphasis from empirical to pre-emptive therapy, especially when substantiated by suggestive radiological findings. These new tools may be used to confirm a presumed diagnosis of invasive aspergillosis, or, when used to screen high-risk patients, may identify an infection at the early stage of disease. The excellent negative predictive value of these assays should convince clinicians to withhold antifungal therapy in persistently febrile neutropenic patients with no other signs of fungal infection. On the other hand, consecutive positive results in a high-risk population should at least trigger a complete diagnostic work-up. This review will focus on the diagnostic utility as well as on the pitfalls of serial screening for the presence of circulating fungal antigens in haematology patients.
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Affiliation(s)
- Johan Maertens
- Department of Haematology, Universitaire Ziekenhuizen Leuven, Catholic University, Leuven, Belgium.
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