title: Simple and rapid detection of swine hepatitis E virus by reverse transcription loop-mediated isothermal amplification.
abstract: 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.
authors.affiliation: Key Laboratory of Animal Disease Control and Prevention of the Ministry of Agriculture, College of Veterinary Medicine, South China Agricultural University, Tianhe District, Guangzhou 510642, People's Republic of China.
authors.forename: Liang-Quan
authors.initials: LQ
authors.lastname: Zhang
authors.forename: Fu-Rong
authors.initials: FR
authors.lastname: Zhao
authors.forename: Zhi-Gang
authors.initials: ZG
authors.lastname: Liu
authors.forename: Wei-Li
authors.initials: WL
authors.lastname: Kong
authors.forename: Heng
authors.initials: H
authors.lastname: Wang
authors.forename: Yun
authors.initials: Y
authors.lastname: Ouyang
authors.forename: Huan-Bin
authors.initials: HB
authors.lastname: Liang
authors.forename: Chao-Yi
authors.initials: CY
authors.lastname: Zhang
authors.forename: Hai-Tao
authors.initials: HT
authors.lastname: Qi
authors.forename: Chang-Li
authors.initials: CL
authors.lastname: Huang
authors.forename: Si-Hu
authors.initials: SH
authors.lastname: Guo
authors.forename: Gui-Hong
authors.initials: GH
authors.lastname: Zhang
mesh_terms: D000818:Animals; D001646:Bile / Q000821:virology; D015967:Gene Expression Regulation, Viral; D016751:Hepatitis E / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D016752:Hepatitis E virus / Q000235:genetics / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods / Q000662:veterinary*; D010802:Phylogeny; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000821:virology*
chemical_list: D012367:RNA, Viral