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Arch Virol ; 157(12): 2383-8, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22855125

RESUMO

Hepatitis E virus (HEV) is an enteric pathogen of humans and animals, and pigs have been considered an important reservoir of this virus. Recent evidence has indicated the cross-species transmission of hepatitis E virus (HEV) from pigs to humans, causing zoonosis, mostly via consumption of uncooked or undercooked animal meat/viscera. In this study, we have developed a one-step RT-LAMP assay for rapid detection of swine HEV. Specific primer sets targeting the ORF3 gene were designed. The sensitivity of the RT-LAMP assay was 10(1) copies/µl of RNA template, which was tenfold higher than that of RT-nPCR. The specificity of this assay was demonstrated by the lack of amplification of DNA/RNA from other swine viruses. Furthermore, a total of 41 bile samples were subjected to RT-LAMP and RT-nPCR. Eighteen positive samples were detected by RT-nPCR, while 36 positive samples were detected by RT-LAMP, indicating that the sensitivity of the RT-LAMP assay was higher than that of the conventional RT-nPCR assay. The RT-LAMP assay reported here may be used for diagnosis of swine HEV, not only in laboratories but also under field conditions.


Assuntos
Vírus da Hepatite E/isolamento & purificação , Hepatite E/veterinária , Técnicas de Amplificação de Ácido Nucleico/veterinária , Doenças dos Suínos/virologia , Animais , Bile/virologia , Regulação Viral da Expressão Gênica , Hepatite E/diagnóstico , Hepatite E/virologia , Vírus da Hepatite E/genética , Técnicas de Amplificação de Ácido Nucleico/métodos , Filogenia , RNA Viral/genética , RNA Viral/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Sensibilidade e Especificidade , Suínos , Doenças dos Suínos/diagnóstico
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