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Langbach O, Kristoffersen AK, Abesha-Belay E, Enersen M, Røkke O, Olsen I. Oral, intestinal, and skin bacteria in ventral hernia mesh implants. J Oral Microbiol 2016; 8:31854. [PMID: 27476443 PMCID: PMC4967714 DOI: 10.3402/jom.v8.31854] [Citation(s) in RCA: 17] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/06/2016] [Revised: 05/29/2016] [Accepted: 06/21/2016] [Indexed: 11/14/2022] Open
Abstract
Background In ventral hernia surgery, mesh implants are used to reduce recurrence. Infection after mesh implantation can be a problem and rates around 6–10% have been reported. Bacterial colonization of mesh implants in patients without clinical signs of infection has not been thoroughly investigated. Molecular techniques have proven effective in demonstrating bacterial diversity in various environments and are able to identify bacteria on a gene-specific level. Objective The purpose of this study was to detect bacterial biofilm in mesh implants, analyze its bacterial diversity, and look for possible resemblance with bacterial biofilm from the periodontal pocket. Methods Thirty patients referred to our hospital for recurrence after former ventral hernia mesh repair, were examined for periodontitis in advance of new surgical hernia repair. Oral examination included periapical radiographs, periodontal probing, and subgingival plaque collection. A piece of mesh (1×1 cm) from the abdominal wall was harvested during the new surgical hernia repair and analyzed for bacteria by PCR and 16S rRNA gene sequencing. From patients with positive PCR mesh samples, subgingival plaque samples were analyzed with the same techniques. Results A great variety of taxa were detected in 20 (66.7%) mesh samples, including typical oral commensals and periodontopathogens, enterics, and skin bacteria. Mesh and periodontal bacteria were further analyzed for similarity in 16S rRNA gene sequences. In 17 sequences, the level of resemblance between mesh and subgingival bacterial colonization was 98–100% suggesting, but not proving, a transfer of oral bacteria to the mesh. Conclusion The results show great bacterial diversity on mesh implants from the anterior abdominal wall including oral commensals and periodontopathogens. Mesh can be reached by bacteria in several ways including hematogenous spread from an oral site. However, other sites such as gut and skin may also serve as sources for the mesh biofilm.
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Affiliation(s)
- Odd Langbach
- Department of Gastroenterologic Surgery, Akershus University Hospital, University of Oslo, Lørenskog, Norway.,Faculty of Medicine, University of Oslo, Oslo, Norway;
| | | | - Emnet Abesha-Belay
- Department of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, Norway
| | - Morten Enersen
- Department of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, Norway
| | - Ola Røkke
- Department of Gastroenterologic Surgery, Akershus University Hospital, University of Oslo, Lørenskog, Norway.,Faculty of Medicine, University of Oslo, Oslo, Norway
| | - Ingar Olsen
- Department of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, Norway
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The Effect of Curcumin on an Animal Intestinal Ischemia/Reperfusion Model for Bacterial Translocation and Inflammatory Response. Int Surg 2015. [DOI: 10.9738/intsurg-d-15-00004.1] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/17/2022] Open
Abstract
Ischemia/reperfusion (IR) injury of the intestine is a major problem in abdominal pathological condition and is associated with a high morbidity and mortality. The purpose of the study is to investigate the effects of curcumin on the bacterial translocation incidence and inflammatory response in rats submitted to bowel ischemia reperfusion injury. Thirty-two Wistar albino rats with a weight of 200 to 250 g were used in the study. They were randomly divided into 3 groups (n = 10 for each group): sham only operated group(group I); IR group (group II); and IR + curcumin treatment group (group III). Curcumin (curcumin from Curcuma longa) 20 mg/kg/day was given orally to the curcumin group. All animals were given 109 E. Coli by orogastric intubation 12 hours before sampling. Seventy-two hours after the first operation, mesenteric lymph node and blood samples were obtained and cultured. Blood samples of 2 mL were obtained for a polymerase chain reaction study. A piece of terminal ileum was also sampled for histopathologic examination. Mesenteric lymph node and blood cultures of all control animals were positive for microbiological growth, and polymerase chain reaction results were positive in seven of the eight rats. Histopathologically, edema, vasodilatation and inflammatory cell infiltration were found to be less in the other groups in comparison to the control group. Curcumin reduced bacterial translocation in blood, hepatocellular damage, and plasma cytokine levels. Curcumin reduced the incidence of bacterial translocation in intestinal I/R. rats. These results suggest that Curcumin would be clinically useful in the treatment of intestinal I/R injury.
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Topuz Ö, Ilhan YS, Doğru O, Aygen E, Sözen S. Effect of melatonin and misoprostol on bacterial translocation in portal hypertensive rats. J Gastroenterol Hepatol 2012; 27:562-5. [PMID: 21793915 DOI: 10.1111/j.1440-1746.2011.06875.x] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/11/2022]
Abstract
BACKGROUND AND AIM Portal hypertension is the main complication of cirrhosis and it is responsible for its most common complications. Bacterial translocation increases the morbidity and mortality rates in patients with portal hypertension. We aimed to investigate the effects of melatonin and misoprostol on bacterial translocation induced by portal hypertension. METHODS We established four groups, each containing eight rats. Except for the control and sham groups, the animals in the other groups (treatment groups) received misoprostol or melatonin for 3 days after the first operation. In the sham group, a laparotomy was carried out and only the portal vein was dissected. Calibrated portal vein ligation was carried out in the other groups. All animals were given 10(10) Escherichia coli by orogastric intubation 12 h before sampling. Seventy-two hours after the first operation, mesenteric lymph node and blood samples were obtained and cultured. Two cc blood samples were obtained for a polymerase chain reaction study. A piece of terminal ileum was also sampled for histopathologic examination. RESULTS Mesenteric lymph node and blood cultures of all control animals were positive for microbiological growth, and polymerase chain reaction results were positive in seven of the eight rats. Histopathologically, edema, vasodilatation and inflammatory cell infiltration were found to be less in the other groups in comparison to the control group. The incidence of bacterial translocation was decreased in all treatment groups as compared to the control group. CONCLUSIONS In this study, bacterial translocation occurred in portal hypertension. Melatonin and misoprostol reduced the incidence of bacterial translocation in portal hypertensive rats.
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Affiliation(s)
- Ömer Topuz
- Department of General Surgery, Kayseri Training and Research Hospital, Kayseri, Turkey.
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Escherichia coli translocation in experimental short bowel syndrome: probiotic supplementation and detection by polymerase chain reaction. Pediatr Surg Int 2011; 27:1301-5. [PMID: 21748652 DOI: 10.1007/s00383-011-2943-z] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Accepted: 10/29/2009] [Indexed: 10/18/2022]
Abstract
BACKGROUND After massive bowel resection, bacterial overgrowth is frequent and favors the occurrence of Gram-negative intestinal bacterial translocation (BT). Probiotics have been recommended in several diseases and may also have beneficial effects on BT. Conversely, polymerase chain reaction (PCR) technique has shown better sensitivity than conventional methods in bacterial detection and has not been investigated in experimental models of short bowel syndrome and BT. OBJECTIVE To test the hypothesis that Bifidobacterium lactis (BL) administration decreases Escherichia coli bacterial translocation (ECBT) in experimental short bowel syndrome and to confirm the better sensitivity of PCR technique to detect ECBT. METHODS Forty-eight adult Wistar rats, orally fed with standard rat chow and tap water ad libitum, were maintained in individual metabolic cages for 10 days and divided into three groups: Control group (n = 15): non-manipulated animals. RES group (n = 15): 80% gut resection. 1 ml of sterile water was administered daily after orogastric intubation. RES-PRO group (n = 18): same resection as RES group and daily administration of 7.8 × 10(9) BL (CFU). At the end of the study, portal blood, peripheral blood and mesenteric lymph node (MLN) samples were recovered and cultured. Also, genomic DNA from E. coli was detected by PCR technique. RESULTS In conventional culture there was no ECBT in control animals whereas 73% of RES and 33% of RES-PRO animals showed it. PCR detected ECBT in 47, 87 and 33%, respectively, showing higher sensitivity.
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Intraoperative Bacterial Translocation Detected by Bacterium-Specific Ribosomal RNA-Targeted Reverse-Transcriptase Polymerase Chain Reaction for the Mesenteric Lymph Node Strongly Predicts Postoperative Infectious Complications After Major Hepatectomy for Biliary Malignancies. Ann Surg 2010; 252:1013-9. [DOI: 10.1097/sla.0b013e3181f3f355] [Citation(s) in RCA: 40] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/27/2023]
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Prospective comparison of eubacterial PCR and measurement of procalcitonin levels with blood culture for diagnosing septicemia in intensive care unit patients. J Clin Microbiol 2009; 47:2964-9. [PMID: 19641068 DOI: 10.1128/jcm.00418-07] [Citation(s) in RCA: 18] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/28/2022] Open
Abstract
Rapid identification of infection has a major impact on the clinical course, management, and outcome of critically ill intensive care unit (ICU) patients. We compared the results of PCR and procalcitonin with blood culture for ICU patients suspected of having septicemia. Ninety patients (60 patients meeting the criteria for sepsis and 30 patients not meeting the criteria for sepsis) were evaluated. Compared with blood culture as the gold standard, the sensitivity, specificity, and positive and negative predictive values for PCR were 100%, 43.33%, 46.87%, and 100%, respectively, and for procalcitonin were 100%, 61.66%, 56.6%, and 100%, respectively. The average times required to produce a final result were as follows: PCR, 10 h; blood culture, 33 h; procalcitonin, 45 min. Both PCR and procalcitonin may be useful as rapid tests for detecting septicemia but compared with blood cultures lacked specificity.
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Pearce CB, Zinkevich V, Beech I, Funjika V, Ruiz AG, Aladawi A, Duncan HD. Using the polymerase chain reaction coupled with denaturing gradient gel electrophoresis to investigate the association between bacterial translocation and systemic inflammatory response syndrome in predicted acute severe pancreatitis. World J Gastroenterol 2006; 11:7142-7. [PMID: 16437661 PMCID: PMC4725083 DOI: 10.3748/wjg.v11.i45.7142] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
AIM To investigate the use of PCR and DGGE to investigate the association between bacterial translocation and systemic inflammatory response syndrome in predicted severe AP. METHODS Patients with biochemical and clinical evidence of acute pancreatitis and an APACHE II score > or = 8 were enrolled. PCR and DGGE were employed to detect bacterial translocation in blood samples collected on d 1, 3, and 8 after the admission. Standard microbial blood cultures were taken when there was clinical evidence of sepsis or when felt to be clinically indicated by the supervising team. RESULTS Six patients were included. Of all the patients investigated, only one developed septic complications; the others had uneventful illness. Bacteria were detected using PCR in 4 of the 17 collected blood samples. The patient with sepsis was PCR-positive in two samples (taken on d 1 and 3), despite three negative blood cultures. Using DGGE and specific primers, the bacteria in all blood specimens which tested positive for the presence of bacterial DNA were identified as E coli. CONCLUSION Our study confirmed that unlike traditional microbiological techniques, PCR can detect the presence of bacteria in the blood of patients with severe AP. Therefore, this latter method in conjunction with DGGE is potentially an extremely useful tool in predicting septic morbidity and evaluating patients with the disease. Further research using increased numbers of patients, in particular those patients with necrosis and sepsis, is required to assess the reliability of PCR and DGGE in the rapid diagnosis of infection in AP.
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Affiliation(s)
- Callum B Pearce
- Gastroenterology, Diagnostic Procedures Unit, Fremantle Hospital, Fremantle, Western Australia, WA 6011, Australia.
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Quigley EMM, Quera R. Small intestinal bacterial overgrowth: roles of antibiotics, prebiotics, and probiotics. Gastroenterology 2006; 130:S78-90. [PMID: 16473077 DOI: 10.1053/j.gastro.2005.11.046] [Citation(s) in RCA: 206] [Impact Index Per Article: 10.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 09/17/2005] [Accepted: 11/14/2005] [Indexed: 12/15/2022]
Abstract
Small intestinal bacterial overgrowth is common in intestinal failure. Its occurrence relates to alterations in intestinal anatomy, motility, and gastric acid secretion. Its presence may contribute to symptoms, mucosal injury, and malnutrition. Relationships between bacterial overgrowth and systemic sepsis are of potential importance in the intestinal failure patient because the direct translocation of bacteria across the intestinal epithelium may contribute to systemic sepsis: a phenomenon that has been well established in experimental animal models. The accurate diagnosis of bacterial overgrowth continues to present a number of challenges in clinical practice and especially so among patients with intestinal failure. The management of patients with bacterial overgrowth remains, for the most part, primarily empiric and comprises antibiotic therapy and correction of any associated nutritional deficiencies. Although evidence from experimental animal studies consistently indicates that probiotics exert barrier-enhancing, antibacterial, immune-modulating, and anti-inflammatory effects, which all could be benefits in small intestinal bacterial overgrowth and intestinal failure, their role in human beings remains to be evaluated adequately.
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Affiliation(s)
- Eamonn M M Quigley
- Alimentary Pharmabiotic Centre, Department of Medicine, National University of Ireland, Cork, Ireland.
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Ono S, Tsujimoto H, Yamauchi A, Hiraki S, Takayama E, Mochizuki H. Detection of microbial DNA in the blood of surgical patients for diagnosing bacterial translocation. World J Surg 2005; 29:535-9. [PMID: 15776295 DOI: 10.1007/s00268-004-7618-7] [Citation(s) in RCA: 20] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/25/2022]
Abstract
Bacterial translocation sometimes occurs in patients during surgical stress and is associated with an increased incidence of septic morbidity. However, no reliable method has been established for diagnosing bacterial translocation in humans. Identification of minute quantities of microbial-specific DNA has been made possible using polymerase chain reaction (PCR) techniques. The aims of this study were to determine the prevalence of bacterial translocation in patients with surgical stress using PCR techniques and to evaluate the usefulness of blood PCR techniques for diagnosing bacterial translocation. DNA was extracted from the blood of 52 surgical patients (24 elective major surgery patients and 28 septic patients) and 10 healthy controls. PCR techniques were used to amplify genes from Escherichia coli, Bacteroides fragilis, a region of 16S ribosomal RNA found in many gram-positive and gram-negative bacteria, and Candida albicans. Bacterial and Candida albicans DNA were not detected in healthy volunteers. Enteric bacterial DNA was detected in patients with hepatic lobectomy, and Candida albicans DNA was detected in patients with esophagectomy on the first postoperative day. Enteric bacterial and Candida albicans DNA were detected in septic patients with findings diagnostic of bacterial translocation, such as small bowel obstruction, ulcerative colitis, or supramesenteric arterial occlusion or in those who had undergone chemotherapy for advanced colon cancer. However, none of the patients were positive by the blood culture technique. The PCR method is more sensitive than blood cultures for detecting bacterial components in the blood of septic patients and is a valuable tool for verifying bacterial translocation in patients who have undergone hepatic lobectomy or esophagectomy. It is also valuable in septic patients who do not have a defined focus of infection.
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Affiliation(s)
- Satoshi Ono
- Department of Surgery I, National Defense Medical College, Namiki 3-2, Tokorozawa, Saitama, 359-8513, Japan.
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Gunji H, Scarth S, Carlson GL, Warhurst G, Little RA, Hopkins SJ. Variability of bacterial translocation in the absence of intestinal mucosal damage following injury and the influence of interleukin-6. ACTA ACUST UNITED AC 2005; 13:39-49. [PMID: 16099144 DOI: 10.1016/j.pathophys.2005.07.002] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/31/2004] [Revised: 07/07/2005] [Accepted: 07/08/2005] [Indexed: 12/22/2022]
Abstract
Bacterial translocation and intestinal mucosal damage have been reported as potentially clinically important sequelae of injury. Evidence that endogenous interleukin-6 (IL-6) is able to protect against infection, and that orally administered IL-6 could prevent bacterial translocation and mucosal damage following haemorrhage, led us to evaluate the impact of injury on the intestinal mucosa and the role of endogenous IL-6. Normal and IL-6-deficient (IL-6-/-) mice were subjected to haemorrhage of increasing severity, hind limb ischaemia, or both. Mucosal integrity and bacterial translocation to the liver, spleen and mesenteric lymph nodes (MLN) were examined after 16 h. Bacterial translocation to each of these tissues was observed in unoperated animals. The more severe haemorrhage procedures, and hind limb ischaemia, increased bacterial translocation to the liver significantly in most experiments with normal mice. The IL-6-/- mice survived the most severe haemorrhage procedure less well (p = 0.0015), although increased bacterial translocation was not seen. There was no clear evidence of mucosal damage, or bacterial translocation to spleen and mesenteric lymph nodes, in either normal or IL-6-/- mice. Intestinal IgA concentrations were the same in IL-6-/- mice and controls. These data demonstrate that increased bacterial translocation can be observed following severe injury, but that neither bacterial translocation nor severe injury are inevitably associated with morphological damage to the intestinal mucosa, and endogenous IL-6 is more likely to promote bacterial translocation than protect the gut.
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Affiliation(s)
- Hirobumi Gunji
- Injury Research Group, Clinical Sciences Building, Hope Hospital, Stott Lane, Salford, Greater Manchester M6 8HD, UK
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Affiliation(s)
- Reiner Wiest
- Department of Internal Medicine I, University Hospital Regensburg, Germany
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João SA, Alencar SSSD, Medeiros ADC, Diniz SOF, Cardoso VN, Brandt CT. Translocation of 99mTc labelled bacteria after intestinal ischemia and reperfusion. Acta Cir Bras 2004. [DOI: 10.1590/s0102-86502004000400003] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/21/2022] Open
Abstract
PURPOSE: Ischemia and reperfusion of the small intestine disrupts gut barrier, causes bacterial translocation and activates inflammatory responses. An experimental study was planned to evaluate if 99mTc labelled Escherichia coli translocates to mesenteric lymph nodes, liver, spleen, lung and serum of rats submitted to mesenteric ischemia/reperfusion. Additionally, it was observed if the time of reperfusion influences the level of translocation. METHODS: Forty male Wistar rats underwent 45 minutes of gut ischemia by occlusion of the superior mesenteric artery. The translocation of labelled bacteria to different organs and portal serum was determined in rats reperfused for 30 minutes, 24 hours, sham(S) and controls(C), using radioactivity count and colony forming units/g (CFU). RESULTS: All the organs from rats observed for 24 hours after reperfusion had higher levels of radioactivity and positive cultures (CFU) than did the organs of rats reperfused for 30 minutes, C and S, except in the spleen (p<0,01). CONCLUSION: The results of this study indicated that intestinal ischemia/reperfusion led to bacterial translocation, mostly after 24 hours of reperfusion.
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Wiest R, Rath HC. Gastrointestinal disorders of the critically ill. Bacterial translocation in the gut. Best Pract Res Clin Gastroenterol 2003; 17:397-425. [PMID: 12763504 DOI: 10.1016/s1521-6918(03)00024-6] [Citation(s) in RCA: 133] [Impact Index Per Article: 6.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 01/31/2023]
Abstract
The human gastrointestinal tract is colonized by a dense population of microorganisms, referred to as the bacterial flora. Although the gut provides a functional barrier between these organisms and the host, bacterial translocation is a common event in the healthy person. However, in critically ill patients, with various underlying diseases, this bacterial translocation may lead to infections and consequently to a further reduction in general health status. The mechanism of bacterial translocation is widely, and somehow controversially investigated in vitro and in animal models. In human studies, several diseases have been associated with bacterial translocation. However, methodological shortcomings, insufficient populations and conflicting results leave many open questions. This is also reflected in the various published therapeutic strategies. To overcome this problem more investigations in humans are needed, especially in techniques for detecting bacterial translocation.
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Affiliation(s)
- Reiner Wiest
- Department of Internal Medicine, University of Regensburg, 93042 Regensburg, Germany.
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Abstract
INTRODUCTION The pathogenesis of acute pancreatitis remains elusive. Sepsis and multiple organ failure continue to cause death (overall mortality rate, approximately 10%) despite immense improvements in supportive, radiologic, and surgical therapy. The gut appears to play a key role in the development of these complications. AIM To critically review the evidence implicating the gut in the pathogenesis of acute pancreatitis. METHODS Relevant English-language literature or abstracts cited in the MEDLINE database were reviewed. RESULTS AND CONCLUSION Gram-negative enteric organisms account for most infections of pancreatic necrosis and subsequent sepsis, which suggests the gut as a source. Intestinal permeability is increased early in patients with severe acute pancreatitis and correlates with endotoxemia, which suggests translocation as a possible mechanism. The pathogenesis of the deranged function of the gut mucosal barrier and the possible sites of increase in intestinal permeability are discussed. The gut also plays a role in priming neutrophils and the release of inflammatory cytokines, which initiate and propagate nearly all the detrimental consequences of severe inflammation and sepsis. Future research avenues and potential therapeutic measures that may restore and preserve gut barrier function are explored.
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Affiliation(s)
- Basil J Ammori
- Division of Surgery, The University of Leeds, and the Center for Digestive Diseases, The General Infirmary, Leeds, United Kingdom.
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Ammori BJ, Fitzgerald P, Hawkey P, McMahon MJ. The early increase in intestinal permeability and systemic endotoxin exposure in patients with severe acute pancreatitis is not associated with systemic bacterial translocation: molecular investigation of microbial DNA in the blood. Pancreas 2003; 26:18-22. [PMID: 12499912 DOI: 10.1097/00006676-200301000-00004] [Citation(s) in RCA: 76] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/11/2022]
Abstract
INTRODUCTION Sepsis is the main cause of late mortality in patients with severe acute pancreatitis and is largely attributed to secondary infection of pancreatic necrosis with gram-negative enteric organisms. This is commonly preceded by a significant increase in intestinal colonization with such microbes and with early increases in intestinal permeability, thus suggesting a mechanism of bacterial translocation. Whilst cultures of blood specimens from these patients often remain sterile, it is conceivable that bacteria might translocate in small volumes with detrimental effects but elude detection by standard microbial culture techniques. AIMS To investigate the incidence and frequency with which bacterial DNA may exist in the systemic circulation of patients with acute pancreatitis and to relate that to disease severity, changes in intestinal permeability, and systemic endotoxin exposure. METHODOLOGY Blood samples were obtained at admission and on days 3 and 7 from 26 patients with acute pancreatitis (seven with severe cases) and from 10 healthy controls for DNA extraction and standard microbial cultures. Polymerase chain reaction techniques were used to amplify a gene region (16S ribosomal RNA) found in all bacteria. Levels of serum endotoxin and antibodies to endotoxin core (EndoCAb) were measured at admission, and intestinal permeability to the macromolecule polyethylene glycol 3350 was determined within 72 hours of the onset of symptoms. RESULTS Blood cultures yielded and enterococci for one patient with a severe attack and coagulase-negative staphylococci for another patient with a mild attack. No bacterial DNA was found in any of the samples. Endotoxemia was detected in 20 patients (five with severe cases), and levels of serum IgM EndoCAb were depleted in patients with severe attacks but remained relatively unchanged during mild attacks (p = 0.033). Intestinal permeability was significantly increased in patients with severe attacks of acute pancreatitis but remained unchanged during mild attacks (p < 0.05). CONCLUSIONS Whilst severe attacks of acute pancreatitis are associated with early derangement in gut barrier function and systemic endotoxin translocation, there is no molecular evidence for associated bacterial "translocation."
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Affiliation(s)
- B J Ammori
- Division of Surgery, The University of Leeds, Leeds, United Kingdom.
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Doty JM, Oda J, Ivatury RR, Blocher CR, Christie GE, Yelon JA, Sugerman HJ. The effects of hemodynamic shock and increased intra-abdominal pressure on bacterial translocation. THE JOURNAL OF TRAUMA 2002; 52:13-7. [PMID: 11791046 DOI: 10.1097/00005373-200201000-00005] [Citation(s) in RCA: 43] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/25/2023]
Abstract
BACKGROUND We hypothesized that hemorrhagic shock followed by the abdominal compartment syndrome (ACS) resulted in bacterial translocation (BT) from the gastrointestinal (GI) tract. METHODS Nineteen Yorkshire swine (20-30 kg) were divided into two groups. In the experimental group, group 1 (n = 10), animals were hemorrhaged to a mean arterial pressure (MAP) of 25-30 mm Hg for a period of 30 minutes and resuscitated to baseline MAP. Subsequently, intra-abdominal pressure (IAP) was increased to 30 mm Hg above baseline by instilling sterile normal saline into the peritoneal cavity. The IAP was maintained at this level for 60 minutes. Acid/base status, gastric mucosal ph (pHi), superior mesenteric artery (SMA) blood flow, and hemodynamic parameters were measured and recorded. Blood samples were analyzed by polymerase chain reaction (PCR) for the presence of bacteria. Spleen, lymph node, and portal venous blood cultures were obtained at 24 hours. Results were analyzed by ANOVA and are reported as mean +/- SEM. The second group was the control. These animals did not have the hemorrhage, resuscitation, or intra-abdominal hypertension (IAH) but were otherwise similar to the experimental group in terms of laparotomy and measured parameters. RESULTS SMA blood flow in group 1 (baseline of 0.87 +/- 0.10 l/min) decreased in response to hemorrhage (0.53 +/- 0.10 l/min, p = 0.0001) and remained decreased with IAH (0.63 l/min +/- 0.10, p = 0.0006) as compared to control and returned towards baseline (1.01 +/- 0.5 l/min) on relief of IAH. pHi (baseline of 7.21 +/- 0.03) was significantly decreased with hemorrhage (7.04 +/- 0.03, p = 0.0003) and decreased further after IAH (6.99 +/- 0.03, p = 0.0001) in group 1 compared to control, but returned toward baseline at 24 hours (7.28 +/- 0.04). The mean arterial pH decreased significantly from 7.43 +/- 0.01 at baseline to 7.27 +/- 0.01 at its nadir within group 1 (p = 0.0001) as well as when compared to control (p = 0.0001). Base excess was also significantly decreased between groups 1 and 2 during hemorrhage (3.30 +/- 0.71 vs. 0.06 +/- 0.60, p = 0.001) and IAH (3.08 +/- 0.71 vs. -1.17 +/- 0.60, p = 0.0001). In group 1, 8 of the 10 animals had positive lymph node cultures, 2 of the 10 had positive spleen cultures, and 2 of the 10 had positive portal venous blood cultures for gram-negative enteric bacteria. Only 2 of the 10 animals had a positive PCR. In group 2, five of the nine animals had positive lymph node cultures, zero of the nine had positive spleen cultures, and one of the nine had positive portal venous blood cultures. Two of the nine animals had positive PCRs. There was no significant difference in cultures or PCR results between the two groups (Fisher's exact test, p = 0.3). CONCLUSION In this study, hemorrhage followed by reperfusion and a subsequent insult of IAH caused significant GI mucosal acidosis, hypoperfusion, as well as systemic acidosis. These changes did not appear to be associated with a significant bacterial translocation as judged by PCR measurements, tissue, or blood cultures.
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Affiliation(s)
- James M Doty
- Department of Surgery, Medical College of Virginia, Virginia Commonwealth University, Richmond, Virginia, USA
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Reno WL, Mcdaniel DO, Turner WW, Williams MD. Polymerase Chain Reaction for the Detection of Bacteremia. Am Surg 2001. [DOI: 10.1177/000313480106700603] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/02/2022]
Abstract
Analysis of blood by polymerase chain reaction (PCR) is a more rapid and sensitive method to detect bacteremia than blood culture. The PCR was performed on blood obtained from patients during blood culture draws and on blood from normal volunteers. Eighty-seven patients provided 125 blood samples for blood culture comparison with PCR. Specific PCR primers for Staphylococcus aureus and Escherichia coli that targeted conserved regions common to Gram-positive and Gram-negative bacteria were used. Selective stringency conditions identified other Gram-positive and Gram-negative bacteria. The blood culture agreed with the PCR in 93 of the 125 patient specimens (74%). In 29 of these specimens the PCR was positive yet the blood culture was negative. When clinical information was included with positive blood culture to define sepsis in these patients and their specimens were added to the positive blood cultures the statistical accuracy of PCR was 93 per cent. Only three of the 78 specimens with negative PCR had positive blood cultures. The PCR was negative in all but one of the 50 volunteers. PCR is more sensitive than blood culture, and it can quickly rule out bacteremia.
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Affiliation(s)
- William L. Reno
- Departments of Surgery, University of Mississippi Medical Center, Jackson, Mississippi
- G.V. (Sonny) Montgomery Veterans Affairs Medical Center, Jackson, Mississippi
| | - D. Olga Mcdaniel
- Departments of Surgery, University of Mississippi Medical Center, Jackson, Mississippi
- Departments of Neurology, University of Mississippi Medical Center, Jackson, Mississippi
| | - William W. Turner
- Departments of Surgery, University of Mississippi Medical Center, Jackson, Mississippi
- G.V. (Sonny) Montgomery Veterans Affairs Medical Center, Jackson, Mississippi
| | - Mark D. Williams
- Departments of Surgery, University of Mississippi Medical Center, Jackson, Mississippi
- G.V. (Sonny) Montgomery Veterans Affairs Medical Center, Jackson, Mississippi
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18
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Zhang WZ, Han TQ, Tang YQ, Zhang SD. Rapid detection of sepsis complicating acute necrotizing pancreatitis using polymerase chain reaction. World J Gastroenterol 2001; 7:289-92. [PMID: 11819777 PMCID: PMC4723539 DOI: 10.3748/wjg.v7.i2.289] [Citation(s) in RCA: 14] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Affiliation(s)
- W Z Zhang
- Department of Surgery, Ruijin Hospital, Shanghai Second Medical University, Shanghai 200025, China
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19
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Schoeffel U, Pelz K, Häring RU, Amberg R, Schandl R, Urbaschek R, von Specht BU, Farthmann EH. Inflammatory consequences of the translocation of bacteria and endotoxin to mesenteric lymph nodes. Am J Surg 2000; 180:65-72. [PMID: 11036145 DOI: 10.1016/s0002-9610(00)00410-4] [Citation(s) in RCA: 17] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/18/2022]
Abstract
BACKGROUND Translocation of intestinal bacteria to mesenteric lymph nodes (MLNs) has been documented in humans under a variety of circumstances, yet its clinical significance remains to be established. The aim of this study was to correlate detectable translocation to MLNs of bacteria and endotoxin with local and systemic signs of inflammation. METHODS From each of 10 patients with carcinoma of the cecal region two MLNs were harvested prior to resection. The presence of bacteria and endotoxin in the lymphatic tissue and blood was determined by culture methods and DNA preparation (PCR) and by a Limulus assay, respectively. Inflammatory mediators were determined in plasma and in MLN homogenates. RESULTS Viable bacteria were detected in MLNs of 7 patients and in 9 of 20 lymph nodes. PCR revealed traces of bacteria in 4 patients and in 6 of their MLNs. Combining both modalities, the translocation rate was 80% and 55% for patients and MLNs, respectively. There was no detectable bacteremia. Endotoxin was found in the plasma of 7 patients and in 9 MLNs from 5 patients. There was no correlation between culture findings and endotoxin concentrations. Moreover, bacteriological data did not correspond to local or systemic inflammation. The group of MLN with detectable endotoxin differed significantly from LPS-negative nodes with respect to interleukin-6, interleukin-10, and sCD14. Systemic concentrations of endotoxin and inflammatory parameters did not correspond to levels within MLNs. CONCLUSION Translocation to MLNs occurs in patients with cecal carcinoma. This, however, seems not to be of major clinical significance if no additional physiologic insults are encountered. Irrespective of the presence of bacteria, there are variations in inflammatory reactions between lymph nodes from one and the same patient, probably reflecting fluctuating response mechanisms to low-grade translocation.
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Affiliation(s)
- U Schoeffel
- Department of Surgery, University of Freiburg, Freiburg, Germany
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20
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Kempf VA, Trebesius K, Autenrieth IB. Fluorescent In situ hybridization allows rapid identification of microorganisms in blood cultures. J Clin Microbiol 2000; 38:830-8. [PMID: 10655393 PMCID: PMC86216 DOI: 10.1128/jcm.38.2.830-838.2000] [Citation(s) in RCA: 276] [Impact Index Per Article: 11.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022] Open
Abstract
Using fluorescent in situ hybridization (FISH) with rRNA-targeted fluorescently labelled oligonucleotide probes, pathogens were rapidly detected and identified in positive blood culture bottles without cultivation and biotyping. In this study, 115 blood cultures with a positive growth index as determined by a continuous-reading automated blood culture system were examined by both conventional laboratory methods and FISH. For this purpose, oligonucleotide probes that allowed identification of approximately 95% of those pathogens typically associated with bacteremia were produced. The sensitivity and specificity of these probes were 100%. From all 115 blood cultures, microorganisms were grown after 1 day and identification to the family, genus, or species level was achieved after 1 to 3 days while 111 samples (96.5%) were similarly identified by FISH within 2.5 h. Staphylococci were identified in 62 of 62 samples, streptococci and enterococci were identified in 19 of 20 samples, gram-negative rods were identified in 28 of 30 samples, and fungi were identified in two of two samples. Thus, FISH is an appropriate method for identification of pathogens grown in blood cultures from septicemic patients.
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Affiliation(s)
- V A Kempf
- Max von Pettenkofer-Institut für Hygiene und Medizinische Mikrobiologie, Ludwig Maximilians Universität München, D-80336 Munich, Germany
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21
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Küçükaydin M, Kocaoğlu C, Köksal F, Kontaş O. Detection of intestinal bacterial translocation in subclinical ischemia-reperfusion using the polymerase chain reaction technique. J Pediatr Surg 2000; 35:41-3. [PMID: 10646771 DOI: 10.1016/s0022-3468(00)80010-x] [Citation(s) in RCA: 14] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 10/25/2022]
Abstract
BACKGROUND/PURPOSE The purpose of this study was to evaluate the detection of bacterial translocation after subclinical ischemia reperfusion injuries in rats with the polymerase chain reaction (PCR) technique. METHODS Six-week-old weaning rats were divided into 3 groups. (1) Experiment rats (n = 20) were gavaged with 10(10) Escherichia coli followed by superior mesentery artery occluded for 10 minutes, then reperfused for 30 minutes. (2) Control rats (n = 20) received bacterial gavage. (3) Group 3 were sham rats (n = 20). After the procedure, 3 mL of blood was obtained from the portal vein. The terminal ileum and mesenteric lymph node (MLN) near the terminal ileum were removed. E. coli DNA was detected in blood and MLN samples by PCR, and histological changes were examined. RESULTS E. coli DNA detection in ischemia-reperfusion (I/R) group animals was 6 of 20 (30%) in the MLN and 2 of 20 (10%) in the blood. PCR was negative in all the rats in the control group and in the sham group (P < .05). There were no significant differences in the histological examination of rat intestines. CONCLUSION These data suggest that subclinical intestinal I/R injury results in bacterial translocation. Also, PCR is a highly sensitive and rapid method to detect the presence of microbial DNA.
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Affiliation(s)
- M Küçükaydin
- Department of Pediatric Surgery, Erciyes University Medical Faculty, Kayseri, Turkey
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22
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Heininger A, Binder M, Schmidt S, Unertl K, Botzenhart K, Döring G. PCR and blood culture for detection of Escherichia coli bacteremia in rats. J Clin Microbiol 1999; 37:2479-82. [PMID: 10405388 PMCID: PMC85261 DOI: 10.1128/jcm.37.8.2479-2482.1999] [Citation(s) in RCA: 75] [Impact Index Per Article: 2.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022] Open
Abstract
Critically ill patients often develop symptoms of sepsis and therefore require microbiological tests for bacteremia that use conventional blood culture (BC) techniques. However, since these patients frequently receive early empirical antibiotic therapy before diagnostic procedures are completed, examination by BC can return false-negative results. We therefore hypothesized that PCR could improve the rate of detection of microbial pathogens over that of BC. To test this hypothesis, male Wistar rats were challenged intravenously with 10(6) CFU of Escherichia coli. Blood was then taken at several time points for detection of E. coli by BC and by PCR with E. coli-specific primers derived from the uidA gene, encoding beta-glucuronidase. In further experiments, cefotaxime (100 or 50 mg/kg of body weight) was administered intravenously to rats 10 min after E. coli challenge. Without this chemotherapy, the E. coli detection rate decreased at 15 min and at 210 min after challenge from 100% to 62% of the animals with PCR and from 100% to 54% of the animals with BC (P, >0.05). Chemotherapy decreased the E. coli detection rate at 25 min and at 55 min after challenge from 100% to 50% with PCR and from 100% to 0% with BC (P, <0.05). Thus, at clinically relevant serum antibiotic levels, PCR affords a significantly higher detection rate than BC in this rat model. The results suggest that PCR could be a useful adjunct tool supplementing conventional BC techniques in diagnosing bacteremia.
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Affiliation(s)
- A Heininger
- Klinik für Anästhesiologie, Hygiene Institute, University of Tübingen, Tübingen, Germany
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23
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DeWitt RC, Kudsk KA. The gut's role in metabolism, mucosal barrier function, and gut immunology. Infect Dis Clin North Am 1999; 13:465-81, x. [PMID: 10340178 DOI: 10.1016/s0891-5520(05)70086-6] [Citation(s) in RCA: 65] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/22/2023]
Abstract
The gastrointestinal tract functions not only to absorb nutrients, it also plays an important immunologic role during health and critical illness. Under experimental and certain clinical conditions, stimulating the gut attentuates the stress response and avoids mucosal atrophy and increases permeability. Gut stimulation prevents atrophy of the gut-associated lymphoid tissue, the body's major defender of moist mucosal surfaces. A better understanding of gut function and improved nutrient delivery has clinical implications in the treatment of critically ill patients.
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Affiliation(s)
- R C DeWitt
- Department of Surgery, University of Tennessee College of Medicine, Memphis, USA
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Sautner T, Wessely C, Riegler M, Sedivy R, Götzinger P, Losert U, Roth E, Jakesz R, Függer R. Early effects of catecholamine therapy on mucosal integrity, intestinal blood flow, and oxygen metabolism in porcine endotoxin shock. Ann Surg 1998; 228:239-48. [PMID: 9712570 PMCID: PMC1191466 DOI: 10.1097/00000658-199808000-00014] [Citation(s) in RCA: 51] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/26/2022]
Abstract
OBJECTIVE To determine the early effects of therapy of endotoxin (ET) shock with epinephrine, norepinephrine, or dopexamine on splanchnic circulation, oxygen metabolism, sigmoid mucosal pHi, bacterial translocation, and morphologic integrity of the ileal, colonic, and sigmoid mucosa. SUMMARY BACKGROUND DATA Conflicting concepts exist concerning the catecholamine therapy of septic shock, but little is known about the effects of catecholamine treatment on splanchnic circulation and mucosal integrity. METHODS ET shock was induced in pigs by ET infusion over 30 minutes, and animals were studied for 4 hours. All animals were resuscitated with fluid. To mimic the treatment of septic shock in humans, mean arterial pressure was maintained in two groups at >70 mm Hg with the administration of epinephrine or norepinephrine. A third group of animals received dopexamine at 7 microg/kg per minute. Systemic and splanchnic blood flow and oxygen metabolism were studied, sigmoid colon mucosal pHi was obtained tonometrically, and bacterial translocation was determined by culture of portal venous blood, mesenteric lymph nodes, liver, spleen, and lung specimens. Histologic sections of ileal, colonic, and sigmoid mucosa were morphometrically examined for therapy effects. RESULTS All investigated catecholamines increased cardiac output and systemic oxygen delivery, whereas intestinal blood flow and oxygen delivery remained unchanged. Sigmoid mucosal pHi decreased in all study animals, but the decrease was most pronounced in the epinephrine group. Pigs receiving epinephrine also showed >40% damage of the mucosa of the ileum and colon, whereas animals receiving ET alone, norepinephrine, or dopexamine showed only moderate lesions with signs of restitution. No animal showed bacterial translocation. CONCLUSIONS Systemic hemodynamics and oxygen metabolism data do not reflect intestinal perfusion. Norepinephrine or dopexamine administration in ET shock causes no additional impairment of intestinal integrity. Epinephrine therapy, in contrast, is associated with a significant reduction of mucosal pHi and considerable early mucosal damage. Its application in septic shock is hazardous.
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Affiliation(s)
- T Sautner
- Department of Surgery, Medical Faculty, University of Vienna, Austria
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Wilmore DW. Polymerase chain reaction surveillance of microbial DNA in critically ill patients: exploring another new frontier. Ann Surg 1998; 227:10-1. [PMID: 9445104 PMCID: PMC1191166 DOI: 10.1097/00000658-199801000-00002] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/05/2023]
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Kane TD, Alexander JW, Johannigman JA. The detection of microbial DNA in the blood: a sensitive method for diagnosing bacteremia and/or bacterial translocation in surgical patients. Ann Surg 1998; 227:1-9. [PMID: 9445103 PMCID: PMC1191165 DOI: 10.1097/00000658-199801000-00001] [Citation(s) in RCA: 96] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/05/2023]
Abstract
OBJECTIVE The purpose was to determine the sensitivity of detecting microbial DNA in the blood of surgical patients as a measure for diagnosing systemic infection and/or translocation from the gut. SUMMARY BACKGROUND DATA Microbial infections and translocation of intestinal bacteria are thought to contribute to multiple system organ failure, but bacterial cultures are often negative in patients with this complication. METHODS DNA was extracted from the blood of 40 surgical patients and 20 healthy controls. Polymerase chain reaction (PCR) techniques were used to amplify genes from Escherichia coli, Bacteroides fragilis, and a region of 16S ribosomal RNA found in many gram-positive and -negative bacteria. RESULTS Bacterial DNA genes were not detected in healthy volunteers but were found in all patients with positive blood cultures. All eight transplant patients receiving OKT3 therapy had microbial DNA in their blood, possibly indicating translocation from the gut. Sixty-four percent of critically ill patients had microbial DNA detected in their blood, but only 3 (14%) had positive blood cultures. CONCLUSIONS The PCR method is more sensitive than blood cultures for detecting bacterial components in the blood of critically ill surgical patients and may detect microbial translocation from the intestine.
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Affiliation(s)
- T D Kane
- Department of Surgery, University of Cincinnati, and the Shriners Burns Institute, Ohio 45267-0558, USA
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