Trends in Reflexive Hepatitis C Virus Testing in a National US Laboratory: A Retrospective Study From 2018 to 2024

Clin Infect Dis. 2026 Jul 8;82(6):988-992. doi: 10.1093/cid/ciag227.

Abstract

Background: In 2019 and 2020, universal hepatitis C screening of adults was recommended by clinical guideline authorities, including the Centers for Disease Control and Prevention (CDC). In 2023, CDC recommendations emphasized the importance of reflexive hepatitis C testing in which samples positive for hepatitis C virus (HCV) antibody (Ab) automatically reflex to viral RNA nucleic acid amplification testing (NAAT). Laboratory stewardship initiatives to guide compliance with expanded guidelines were implemented.

Methods: We conducted a retrospective review of 38 233 788 specimens with HCV Ab testing and 1 423 543 follow-up HCV RNA NAAT performed from 2018 to 2024 to evaluate the impact of expanded hepatitis C guidelines and laboratory stewardship initiatives.

Results: From 2018 to 2024, reflexive hepatitis C testing volumes increased annually, while orders for standalone HCV Ab testing and HCV Ab positivity rates steadily declined. Among HCV Ab-positive individuals, the percentage with reflexive testing increased from 21.9% in 2018 to 82.2% in 2024 (Ƭ = 1.00, P = .003) and RNA testing rates increased from 66.4% in 2018 to 89.3% in 2024 (Ƭ = 1.00, P = .003). Follow-up RNA testing was increasingly completed earlier in HCV Ab-positive individuals with the same-day RNA testing rate increasing from 38.0% in 2018 to 82.2% in 2024. HCV RNA positivity decreased 19.2% during the study period.

Conclusions: Expanded hepatitis C screening guidelines highlighting the importance of reflexive HCV testing and subsequent internal laboratory stewardship initiatives likely contributed to the increased utilization of reflexive hepatitis C testing and the decreased time to identification of active hepatitis C infection.

Keywords: and HCV antibody testing; hepatitis C virus (HCV); reflexive HCV testing; universal HCV screening.

MeSH terms

  • Centers for Disease Control and Prevention, U.S.
  • Hepacivirus* / genetics
  • Hepacivirus* / immunology
  • Hepacivirus* / isolation & purification
  • Hepatitis C Antibodies / blood
  • Hepatitis C* / diagnosis
  • Hepatitis C* / epidemiology
  • Humans
  • Mass Screening* / methods
  • Mass Screening* / trends
  • Nucleic Acid Amplification Techniques
  • RNA, Viral
  • Retrospective Studies
  • United States / epidemiology

Substances

  • Hepatitis C Antibodies
  • RNA, Viral

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