title: Detection of hepatitis E virus RNA in raw sausages and liver sausages from retail in Germany using an optimized method.
abstract: Hepatitis E virus (HEV) is a pathogen of increasing importance, which can be zoonotically transmitted from domestic pigs, wild boar, and deer to humans. Foodborne transmission by consumption of raw and undercooked liver, meat, or sausages prepared from infected animals has been documented. The aim of this study was to investigate the distribution of HEV in different types of sausages sold in Germany. As no standardized methods for HEV detection in food exist, several techniques of sample homogenization, virus concentration and nucleic acid extraction followed by real-time RT-PCR were compared using artificially contaminated sausages. A method using TRI Reagent® Solution showed the best efficacy of matrix disruption and a treatment with chloroform followed by a silica-based RNA extraction method resulted in the highest HEV detection rates. The detection limit of the method was 2.9 × 10(3) and 5.3 × 10(4) genome equivalents per 5 g raw sausage and 2 g liver sausage, respectively. Application of the method to raw and liver sausages from retail in Germany resulted in the HEV genome detection in 14 out of 70 (20%) raw sausages and in 11 out of 50 (22%) liver sausages. The detected HEV sequences showed a high diversity and belonged to different subtypes of HEV genotype 3. The results indicate a broad distribution of HEV-RNA in meat products sold in Germany; however, the infectivity of the detected virus remains to be assessed in future.
authors.affiliation: Institute of Food Quality and Food Safety, University of Veterinary Medicine Hannover, Hannover, Germany,; Unit Food Hygiene and Virology, Federal Institute for Risk Assessment, Berlin, Germany.
authors.forename: Kathrin
authors.initials: K
authors.lastname: Szabo
authors.affiliation: Unit Food Hygiene and Virology, Federal Institute for Risk Assessment, Berlin, Germany. Electronic address: Eva.Trojnar@bfr.bund.de.
authors.forename: Eva
authors.initials: E
authors.lastname: Trojnar
authors.affiliation: Institute of Food Quality and Food Safety, University of Veterinary Medicine Hannover, Hannover, Germany,; Central Institute of the Bundeswehr Medical Service, Kiel, Germany.
authors.forename: Helena
authors.initials: H
authors.lastname: Anheyer-Behmenburg
authors.affiliation: Central Institute of the Bundeswehr Medical Service, Kiel, Germany.
authors.forename: Alfred
authors.initials: A
authors.lastname: Binder
authors.affiliation: Central Institute of the Bundeswehr Medical Service, Kiel, Germany.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Schotte
authors.affiliation: Unit Food Hygiene and Virology, Federal Institute for Risk Assessment, Berlin, Germany.
authors.forename: Lüppo
authors.initials: L
authors.lastname: Ellerbroek
authors.affiliation: Institute of Food Quality and Food Safety, University of Veterinary Medicine Hannover, Hannover, Germany.
authors.forename: Günter
authors.initials: G
authors.lastname: Klein
authors.affiliation: Unit Food Hygiene and Virology, Federal Institute for Risk Assessment, Berlin, Germany.
authors.forename: Reimar
authors.initials: R
authors.lastname: Johne
mesh_terms: D000818:Animals; D001483:Base Sequence; D005506:Food Contamination / Q000032:analysis*; D005517:Foodborne Diseases / Q000517:prevention & control / Q000821:virology; D005858:Germany; D016751:Hepatitis E / Q000517:prevention & control / Q000635:transmission; D016752:Hepatitis E virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D017909:Levivirus / Q000235:genetics / Q000302:isolation & purification; D057230:Limit of Detection; D008099:Liver / Q000821:virology; D008461:Meat Products / Q000506:poisoning / Q000821:virology*; D012367:RNA, Viral / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction; D017423:Sequence Analysis, RNA; D013552:Swine
chemical_list: D012367:RNA, Viral
keywords: Detection method; Hepatitis E virus; Pig liver products; Pork sausages; RT-PCR; Real-time RT-PCR