Author(s)

Zhiwei Sui, Siyuan Liu, Sizhang Liu, Jing Wang, Lei Xue, Xiaoxia Liu, Bin Wang, Shaopeng Gu, and Yi Wang

Corresponding Author

Zhiwei Sui

Abstract

Hepatitis A virus (HAV) infection is the leading worldwide cause of acute viral hepatitis, and outbreaks caused by this virus often occur in contaminated water and food. Foodborne enteric viruses are conventionally processed by quantitative RT-PCR (RT-qPCR), which gives sensitive and relative quantitative detection results based on the standard curve. While RT-qPCR has limitations and may lead to incomparability of results from different laboratories. Here, we developed a reverse transcription digitial PCR (RT-dPCR) for absolute quantification of HAV genomic RNA. Our data showed that RT-dPCR assay achieved highest precision (RSD<6%), when optimum range was between 284 and 2029 copies per panel, especially the lowest RSD was 1.6%, when the number of copies per panel was 1008. In addition, RT-dPCR assays had a lower limit of detection of 2.6 copies/µL HAV genomic RNA, compared with that of approximately 10 copies/µL by RT-qPCR assay. This study demonstrates that RT-dPCR is capable of absolution and accurate quantification of low copy RNA targets of HAV in food products, water samples and vaccine products.

Keywords

Hepatitis A virus, RT-dPCR, absolute quantification, accurate quantification