Published online Aug 6, 2022. doi: 10.5528/wjtm.v10.i2.14
Peer-review started: April 2, 2022
First decision: May 31, 2022
Revised: June 14, 2022
Accepted: July 16, 2022
Article in press: July 16, 2022
Published online: August 6, 2022
Acute ischemic stroke (AIS) is one of the major causes of the continuous increasing rate of global mortality due to the lack of timely diagnosis, prognosis, and management. This study provides a primitive platform for non-invasive and cost-effective diagnosis and prognosis of patients with AIS using circulating cell-free mitochondrial DNA (cf-mtDNA) quantification and validation.
To evaluate the role of cf-mtDNA as s non-invasive, and affordable tool for real-time monitoring and prognosticating AIS patients at disease onset and during treatment.
This study enrolled 88 participants including 44 patients with AIS and 44 healthy controls with almost similar mean age group at stroke onset, and at 24 h and 72 h of treatment. Peripheral blood samples were collected from each study participant and plasma was separated using centrifugation. The cf-mtDNA concentration was quantified using nanodrop reading and validated through real-time quantitative polymerase chain reaction (RT-qPCR) of NADH-ubiquinone oxidoreductase chain 1 (ND1) relative transcript expression levels.
Comparative analysis of cf-mtDNA concentration in patients at disease onset showed significantly increased levels compared to control individuals for both nanodrop reading, as well as ND1 relative expression levels (P < 0.0001). Intergroup analysis of cf-mtDNA concentration using nanodrop showed significantly reduced levels in patients at 72 h of treatment compared to onset (P < 0.01). However, RT-qPCR analysis showed a significant reduction at 24 h and 72 h of treatment compared to the disease onset (P < 0.001). The sensitivity and specificity were relatively higher for RT-qPCR than nanodrop-based cf-mtDNA quantification. Correlation analysis of both cf-mtDNA concentration as well as ND1 relative expression with National Institute of Health Stroke Scale score at baseline showed a positive trend.
In summary, quantitative estimation of highly pure cf-mtDNA provides a simple, highly sensitive and specific, non-invasive, and affordable approach for real-time monitoring and prognosticating AIS patients at onset and during treatment.
Core Tip: Several blood biomarkers have been studied to determine the severity and outcome of ischemic stroke with limited applications. Hence, we need to establish molecular markers which can provide more comprehensive information on the stroke pathophysiology and treatment response. Dynamic quantification of plasma cell-free DNA appears to be a valid and reliable option. Hence, we compared the real-time expression of cell-free mitochondrial DNA (NADH-ubiquinone oxidoreductase chain 1 gene) in ischemic stroke patients with healthy controls and studies its prognostic value during the treatment. This study could aid in the development of clinical values for assessing real-time, non-invasive mode of ischemic stroke status in the future.