First detection of hepatitis E virus in Austrian pigs by RT-qPCR.
Long known to cause disease outbreaks in man in countries with poor sanitary conditions, an increasing number of autochthonous HEV genotype 3 infections have been reported in industrialised countries. Genotype 3 poses an important potential zoonotic threat, with infected pigs functioning as the main reservoir. This study reports the first detected emergence of HEV in Austrian pigs. Five Austrian strains were partially sequenced and phylogenetic analysis demonstrates that they cluster within genotype 3. In addition, a reverse transcription quantitative real-time PCR (RT-qPCR) method using a MGB-hydrolysis probe was developed offering the possibility to detect the HEV genotype 3 in faeces, liquid- and tissue-samples from domestic pigs. The method was adapted to the strains found in Austria. Sensitivity of the assay was tested with different pig organs (liver, mesenteric lymph nodes and kidney) as well as with serum, bile and faeces samples. Within the dynamic range of the assay, a quantitative determination of virus loads was performed. For specificity testing several common swine pathogens were used. Results demonstrated that the proposed method allows implementation of reliable high-throughput screening of Austrian swine samples in the future.
Austrian Agency for Health and Food Safety (AGES), Institute for Veterinary Disease Control, Moedling, Austria. doris.zwettler@ages.at
Doris
D
Zwettler
Maria
M
Fink
Sandra
S
Revilla-Fernändez
Adolf
A
Steinrigl
Petra
P
Winter
Josef
J
Köfer
D000818:Animals; D001317:Austria; D001646:Bile / Q000821:virology; D005243:Feces / Q000821:virology; D005838:Genotype; D016751:Hepatitis E / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D016752:Hepatitis E virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D007668:Kidney / Q000821:virology; D008099:Liver / Q000821:virology; D008198:Lymph Nodes / Q000821:virology; D010802:Phylogeny; D012367:RNA, Viral / Q000097:blood / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D012680:Sensitivity and Specificity; D017423:Sequence Analysis, RNA / Q000662:veterinary; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology*; D019562:Viral Load
D012367:RNA, Viral