Abstract
Hepatitis A virus (HAV) is a serious threat to public health worldwide. We used multiplex polymerase chain reaction (PCR)-based next-generation sequencing (NGS) to derive information on viral genetic diversity and conduct precise phylogenetic analysis. Four HAV genome sequences were obtained using multiplex PCR-based NGS. HAV whole-genome sequence of one sample was obtained by conventional Sanger sequencing. The HAV strains demonstrated a geographic cluster with sub-genotype IA strains in the Republic of Korea. The phylogenetic pattern of HAV viral protein (VP) 3 region showed no phylogenetic conflict between the whole-genome and partial-genome sequences. The VP3 region in serum and stool samples showed sensitive detection of HAV with differences of quantification that did not exceed <10 copies/μL than the consensus VP4 region using quantitative PCR (qPCR). In conclusion, multiplex PCR-based NGS was implemented to define HAV genotypes using nearly whole-genome sequences obtained directly from hepatitis A patients. The VP3 region might be a potential candidate for tracking the genotypic origin of emerging HAV outbreaks. VP3-specific qPCR was developed for the molecular diagnosis of HAV infection. This study may be useful to predict for the disease management and subsequent development of hepatitis A infection at high risk of severe illness.
Highlights
Hepatitis A virus (HAV) infections are the critical etiology of viral hepatitis and impose a massive socioeconomic burden worldwide [1]
The results demonstrated that HAV VP3 gene might be a potential candidate for tracing the genotypic origin of an outbreak, showing a well-established phylogenic pattern compared with the whole-genome sequences of HAV
Our findings provide significant insights into the genomic epidemiology, genetic identification, and molecular diagnosis of hepatitis A infection
Summary
Hepatitis A virus (HAV) infections are the critical etiology of viral hepatitis and impose a massive socioeconomic burden worldwide [1]. HAV is mainly transmitted via the fecaloral route through contaminated food and water. The prevalence rate of HAV in different countries varies with hygiene levels, and approximately, 1.5 million people are annually infected with HAV worldwide. In the late 1970s and early 1990s, 85–95% of the population were serologically positive for anti-HAV immunoglobulin in Republic of Korea (ROK), China, Japan, Philippines, and Thailand [2]. The seroprevalence of anti-HAV antibodies in the Korean population rapidly declined from more than 80% in the 1970s to less than 20%
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