Viruses (Jan 2022)

Development and Evaluation of a Molecular Hepatitis A Virus Assay for Serum and Stool Specimens

  • Robert A. Kozak,
  • Candace Rutherford,
  • Melissa Richard-Greenblatt,
  • N. Y. Elizabeth Chau,
  • Ana Cabrera,
  • Mia Biondi,
  • Jamie Borlang,
  • Jaqueline Day,
  • Carla Osiowy,
  • Sumathi Ramachandran,
  • Nancy Mayer,
  • Laurel Glaser,
  • Marek Smieja

DOI
https://doi.org/10.3390/v14010159
Journal volume & issue
Vol. 14, no. 1
p. 159

Abstract

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Hepatitis A virus (HAV) is an emerging public health concern and there is an urgent need for ways to rapidly identify cases so that outbreaks can be managed effectively. Conventional testing for HAV relies on anti-HAV IgM seropositivity. However, studies estimate that 10–30% of patients may not be diagnosed by serology. Molecular assays that can directly detect viral nucleic acids have the potential to improve diagnosis, which is key to prevent the spread of infections. In this study, we developed a real-time PCR (RT-PCR) assay to detect HAV RNA for the identification of acute HAV infection. Primers were designed to target the conserved 5′-untranslated region (5′-UTR) of HAV, and the assay was optimized on both the Qiagen Rotor-Gene and the BD MAX. We successfully detected HAV from patient serum and stool samples with moderate differences in sensitivity and specificity depending on the platform used. Our results highlight the clinical utility of using a molecular assay to detect HAV from various specimen types that can be implemented in hospitals to assist with diagnostics, treatment and prevention.

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