Related Experiment Videos
Evaluation of western blot in routine diagnosis of hepatitis C virus
M Tsopanomichalou1, M Ergazaki, D A Spandidos
1Laboratory of Clinical Virology, Medical School, University of Crete, Heraklion, Greece.
Insights
Detecting Hepatitis C virus (HCV) infection is crucial for prevention. Routine testing shows inconsistencies, with only 77.4% of dual ELISA-positive samples confirmed, highlighting the need for improved diagnostic accuracy.
Area of Science:
- Hepatology
- Virology
- Clinical Diagnostics
Background:
- Hepatitis C virus (HCV) is a primary cause of parenterally transmitted hepatitis.
- Accurate detection of HCV infection is vital for reducing transmission rates and effective public health prevention strategies.
- Current diagnostic assays for HCV can yield ambiguous or controversial results, complicating patient management and screening.
Purpose of the Study:
- To investigate the discrepancies and challenges encountered during routine Hepatitis C virus (HCV) testing.
- To evaluate the concordance between Enzyme-Linked Immunosorbent Assay (ELISA) and confirmatory tests, including Polymerase Chain Reaction (PCR).
- To identify specific issues, such as Western blot (WB) negative but PCR-positive results, that impact screening for blood and organ donation.
Main Methods:
- Utilized two Enzyme-Linked Immunosorbent Assay (ELISA) kits for initial screening.
- Employed three different confirmatory assays to validate ELISA results.
- Incorporated Polymerase Chain Reaction (PCR) analysis on a subset of samples to detect viral RNA, serving as a molecular benchmark.
Main Results:
- Only 77.4% of samples testing positive by both ELISA methods achieved confirmation.
- Results classified as 'controversial' by ELISA remained inconsistent when subjected to different confirmatory tests.
- Polymerase Chain Reaction (PCR) data indicated that some Western blot (WB) negative samples were positive for the HCV genome, necessitating exclusion from donation screening.
Conclusions:
- Routine HCV diagnostic assays present significant challenges, leading to a substantial proportion of unconfirmed positive results.
- The variability in confirmatory testing underscores the need for standardized and more reliable diagnostic protocols.
- Improvements in routine HCV screening assays are essential to minimize ambiguous results and ensure the safety of blood and organ donations, as PCR is not feasible for routine screening.
Abstract:
Hepatitis C virus (HCV) is among the major causes of parenterally transmitted hepatitis. Detection of infected persons would greatly diminish transmission rates and is therefore a significant parameter for prevention. Current assays are not able to resolve all cases and sometimes the results are controversial. The present study outlines problems that arise during routine testing. Two ELISA tests and three confirmatory tests were used and Polymerase Chain Reaction (PCR) data were available for some of the samples. The results of this study show that only 77.4% of samples positive for both ELISAs were confirmed as being positive. Controversial ELISA results remained controversial, depending on the confirmatory test used. PCR results, though not complete, point to the major problem of Western blot (WB) negative sera that prove positive for the viral genome and have to be excluded from screening for blood and organ donation. Since PCR cannot be used as a routine screening procedure, improvement of the routine assays is needed to minimize ambiguous results.