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Interlaboratory Validation of a Method for Hepatitis E Virus RNA Detection in Meat and Meat Products

Author:
Althof, Nadine, Trojnar, Eva, Böhm, Thomas, Burkhardt, Sabine, Carl, Anja, Contzen, Matthias, Kilwinski, Jochen, Mergemeier, Steffen, Moor, Dominik, Mäde, Dietrich, Johne, Reimar
Source:
Food and environmental virology 2019 v.11 no.1 pp. 1-8
ISSN:
1867-0334
Subject:
Orthohepevirus A, pork, silica, humans, hepatitis E, liver sausage, food matrix, surveys, swine, wild boars, genotype, outbreak investigation, RNA, homogenization, Europe
Abstract:
Increasing numbers of hepatitis E cases are currently recognized in many European countries. The zoonotic hepatitis E virus (HEV) genotype 3 mainly circulates in domestic pigs and wild boars, and can be transmitted to humans via consumption of insufficiently heated meat or meat products produced from those animals. Here, a detailed protocol for detection of HEV RNA in meat products is provided, which is based on the method originally described by Szabo et al. (Intl J Food Microbiol 215:149–156, 2015). It consists of a TRI Reagent®/chloroform-based food matrix homogenization, a silica bead-based RNA extraction and a real-time RT-PCR-based RNA detection. The method was further validated in a ring trial with nine independent laboratories using pork liver sausage samples artificially contaminated with different amounts of HEV. The results indicate sufficient sensitivity, specificity, and accuracy of the method for its broad future use in survey studies, routine food control or outbreak investigations.
Agid:
6310143