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

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 d...

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Bibliographic Details
Published in:	Viruses Vol. 14; no. 1; p. 159
Main Authors:	Kozak, Robert A, Rutherford, Candace, Richard-Greenblatt, Melissa, Chau, N Y Elizabeth, Cabrera, Ana, Biondi, Mia, Borlang, Jamie, Day, Jaqueline, Osiowy, Carla, Ramachandran, Sumathi, Mayer, Nancy, Glaser, Laurel, Smieja, Marek
Format:	Journal Article
Language:	English
Published:	Switzerland MDPI AG 15-01-2022 MDPI
Subjects:	5' Untranslated Regions Antibodies Chicken pox clinical diagnostics Cytomegalovirus Disease Outbreaks DNA Primers Epidemics Feces - virology Genomes Genotype Hepatitis A Hepatitis A - diagnosis Hepatitis A - virology Hepatitis A virus - genetics Hepatitis B Hepatitis C Herpes viruses Humans Immunoglobulin M Infections Laboratories Limit of Detection Molecular Diagnostic Techniques - methods Molecular Diagnostic Techniques - standards Patients Phylogeny Polymerase chain reaction Public health real-time PCR RNA, Viral Sensitivity and Specificity Serology Serum - virology viral hepatitis England Canada United Kingdom > UK United States > US clinical diagnostics viral hepatitis real-time PCR
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Summary:	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.
Bibliography:	ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Undefined-1 ObjectType-Feature-3 content type line 23
ISSN:	1999-4915 1999-4915
DOI:	10.3390/v14010159