A Probe-Based qPCR Assay for Quantification and Classification of Hepatitis E Virus

The hepatitis E virus (HEV) is an important public health concern globally. Detection of viral nucleic acids provides a highly sensitive and specific approach for epidemiological and diagnostic purpose. In the present report, a probe-based qPCR assay for quantitative detection of HEV was developed and applied for genotype characterization of the HEV in clinical samples. The lower quantification limit of DNA was equivalent to 10 GC per reaction, indicating that the sensitivity of the qPCR assay was excellent. All enteroviruse samples detected by our assay (Hepatitis A virus, Rotavirus, EV 71, Norwalk virus and Poliovirus) were negative, showing 100% sensitivity and specificity. In addition, we screened 20 fecal samples collected from hospitalized patients with hepatitis E using the qPCR assay. The results showed that abundance of HEV range from 2.01 x 10(3) to 1.71 x 10(5) GC/g-samples and single product of the HEV- positive samples was amplified by the assay. Sequence analysis showed that the isolates shared 100% identity with isolates from genotype 4, indicating that the assay can be applied for genotype characterization of the HEV in clinical samples.