Observational studyBMC infectious diseases2026
Missed confirmatory HCV RNA testing in anti-HCV-reactive patients and the role of reflex testing in improving completion of the hepatitis C diagnostic care cascade: a laboratory-based retrospective study.
Observational study in BMC infectious diseases, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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3 authors.
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Abstract
backgroundCompletion of confirmatory HCV RNA testing after anti-HCV reactivity is a critical step in the hepatitis C diagnostic care cascade. However, missed confirmatory testing and repeated anti-HCV testing within routine hospital laboratory workflows remain insufficiently characterized. This study aimed to evaluate the frequency of missed confirmatory HCV RNA testing among anti-HCV-reactive patients, investigate the presence of active infection using archived serum samples, and assess workflow-related diagnostic gaps that could potentially be addressed through laboratory-based reflex testing strategies.
methodsThis retrospective laboratory-based observational study included patients tested for anti-HCV antibodies at a tertiary-care university hospital between January 1 and December 31, 2025. Anti-HCV-reactive patients without confirmatory HCV RNA testing were identified, and retrospective HCV RNA testing was performed on available archived serum samples. Repeated anti-HCV testing patterns and clinic-based differences in confirmatory testing requests were also analyzed to evaluate potential diagnostic workflow gaps.
resultsAmong 27,984 anti-HCV tests performed in 23,833 unique patients, the anti-HCV seroprevalence rate was 1.28%. Confirmatory HCV RNA testing was not requested in 32.3% of anti-HCV-reactive patients. Retrospective testing of archived serum samples from 79 patients without initial confirmatory testing identified active infection in 3.8% of cases (3/79). Confirmatory testing rates were significantly higher in specialist clinics than in non-specialist clinics (95.0% vs. 71.0%, p < 0.001). Repeated anti-HCV testing was observed in 13.2% of patients, and no seroconversion was detected among individuals tested six or more times.
conclusionsMissed confirmatory HCV RNA testing represents an important gap in the diagnostic care cascade, particularly in non-specialist clinical settings. Laboratory-based reflex HCV RNA testing strategies may improve completion of confirmatory testing and reduce unnecessary repeat serological testing, supporting more effective diagnostic stewardship and contributing to hepatitis C elimination efforts. CLINICAL TRIAL NUMBER: Not applicable.
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