Multiplex hydrolysis probe real-time PCR for simultaneous detection of hepatitis A virus and hepatitis E virus.

Feng Qiu,Jingyuan Cao,Shengli Bi,Qiudong Su,Yao Yi

doi:10.3390/ijms15069780

Abstract

Detection of hepatitis viral infections has traditionally relied on the circulating antibody test using the enzyme-linked immunosorbent assay. However, multiplex real-time PCR has been increasingly used for a variety of viral nucleic acid detections and has proven to be superior to traditional methods. Hepatitis A virus (HAV) and hepatitis E virus (HEV) are the major causes of acute hepatitis worldwide; both HAV and HEV infection are a main public health problem. In the present study, a one-step multiplex reverse transcriptase quantitative polymerase chain reaction assay using hydrolysis probes was developed for simultaneously detecting HAV and HEV. This novel detection system proved specific to the target viruses, to be highly sensitive and to be applicable to clinical sera samples, making it useful for rapid, accurate and feasible identification of HAV and HEV.

Highlights

Hepatitis A virus (HAV) infection is the leading cause of acute viral hepatitis throughout the world [1]
The Hepatitis E virus (HEV) quantitative standard was prepared in the same way as HAV, whereas the linear regression equation of the HEV standard curve is Y = −3.923X + 45.328 (R2 = 0.999) (Figure 1)
Sera samples ranged from 5.18 × to 4.93 × 107/mL, whereas virus copies obtained from HEV sera samples ranged from 9.33 × to 4.06 × 107/mL. Both HAV and HEV infection are mainly propagated via the fecal-oral route, which is highly associated with a lack of sanitation infrastructure

Summary

Introduction

Hepatitis A virus (HAV) infection is the leading cause of acute viral hepatitis throughout the world [1]. Hepatitis E virus (HEV), the same as HAV, is a small, non-enveloped RNA virus that can cause acute hepatitis in humans [2]. Both viruses are resistant to freezing, detergents, as well as some. The most significant outbreak of HAV infection occurred in Shanghai, China, in 1988, in which almost 300,000 cases were caused by the consumption of clams harvested from a sewage-polluted area [4]. HAV and HEV outbreaks mainly occurred in developing countries where sanitation is not typically available [7]

Methods

Results

Discussion

Conclusion