Comparison of Real-Time Reverse Transcriptase PCR Assays for Detection of Swine Hepatitis E Virus in Fecal Samples

Abstract

dHepatitis E virus (HEV) is a major cause of acute viral hepatitis in people in many developing countries and is also endemic in many industrialized countries. Mammalian HEV (mHEV) isolates can be divided into at least four recognized major genotypes. Several nucleic acid amplification techniques have been developed for mHEV detection, with great differences in sensitivity. The aim of this study was to compare the performances of two singleplex real-time reverse transcriptase (RT) PCR assays for broad detection of all four mHEV genotypes (assays A and B) and two duplex real-time RT-PCR assays for detection and differentiation of mHEV genotypes 3 and 4 (assays C and D). RNAs extracted from 28 fecal samples from pigs experimentally inoculated with HEV genotype 3 and 186 fecal samples from commercial pigs with unknown HEV exposure were tested by all four assays. In experimental samples, HEV RNA was detected in 96.4% (assay A), 39.2% (assay B), 14.2% (assay C), and 0% (assay D) of the samples. In field samples with unknown HEV exposure, HEV RNA was detected in 67.2% (assay A), 36.4% (assay B), 1.1% (assay C), and 0.5% (assay D) of the samples. The assays showed overall poor agreement ( ‫؍‬ 0.19 to 0.03), with differences in detection rates between assays (P < 0.01). Assays A and B, which broadly detect HEV genotypes 1 to 4, had significantly higher detection rates for HEV RNA than the duplex assays C and D, which were both designed to detect and differentiate between HEV genotypes 3 and 4.

Respiratory and gastrointestinal infections at the 2017 Grand Magal de Touba, Senegal: A prospective cohort survey

“…The Multiplex RNA Virus Master Kit (Roche Diagnosis, France) was used for detecting hepatitis A and E virus, adenovirus, rotavirus, norovirus, astrovirus and internal controls MS2 phage by one-step simplex real-time quantitative RT-PCR amplifications [9][10][11].…”

Detection and quantification of hepatitis E virus in the absence of IgG and IgM anti-HEV in HIV-positive patients

“…This fact has been related to low titres of anti‐HEV IgG and IgM antibodies in serum or, probably, due to late or nonexistent seroconversion, as the remains of these antibodies in blood can vary in an undetermined way (Gerber et al . ). Additionally, in accordance with Pischke et al .…”

Effect of swine hepatitis E virus on the livers of experimentally infected Mongolian gerbils by swine hepatitis E virus

“…Following the method described by Priscilla F. Gerber, purified high-quality plasmid DNA was prepared for absolute quantification from the HEV ORF3 region (Gerber et al, 2014). The amount of the recombinant plasmid which was synthesized by the BGI company (China) was quantified using a Scandrop RS232 spectrophotometer according to the manufacturer's instructions (Analytikjena, Germany) and converted into genome copy numbers (Gerber et al, 2014). A standard curve was generated using plasmid DNA, and the HEV titers of the samples were determined based on the standard curve.…”