文献类型：期刊文献

英文题名：Development of a real-time loop-mediated isothermal amplification (LAMP) assay and visual LAMP assay for detection of African swine fever virus (ASFV)

作者：Wang, Deguo[1];Yu, Jianghan[2];Wang, Yongzhen[1];Zhang, Meng[2];Li, Peng[3];Liu, Meng[4];Liu, Yanhong[5]

第一作者：Wang, Deguo

通讯作者：Liu, YH[1]

机构：[1]Xuchang Univ, Key Lab Biomarker Based Rapid Detect Technol Food, Xuchang 461000, Peoples R China;[2]Henan Univ Sci & Technol, Sch Food & Biol Engn, Luoyang 471000, Peoples R China;[3]XinXiang Univ, Xinxiang 453000, Henan, Peoples R China;[4]Zhumadian Vet Drug & Feed Anim Prod Inspect & Tes, Zhumadian 463000, Peoples R China;[5]ARS, Mol Characterizat Foodborne Pathogens Res Unit, Eastern Reg Res Ctr, ARS, Wyndmoor, PA 19038 USA

第一机构：Xuchang Univ, Key Lab Biomarker Based Rapid Detect Technol Food, Xuchang 461000, Peoples R China

通讯机构：[1]corresponding author), ARS, Mol Characterizat Foodborne Pathogens Res Unit, Eastern Reg Res Ctr, ARS, Wyndmoor, PA 19038 USA.

年份：2020

卷号：276

外文期刊名：JOURNAL OF VIROLOGICAL METHODS

收录：;Scopus(收录号：2-s2.0-85074931839);WOS:【SCI-EXPANDED(收录号:WOS:000508749900003)】；

基金：This research was funded by the National Key Research and Development Program of China, grant number 2016YFD0500704-4, and by Henan Science and Technology Plan Project, grant number 182102110285.

语种：英文

外文关键词：African swine fever (ASF); African swine fever virus (ASFV); Loop-mediated isothermal amplification (LAMP)

摘要：African swine fever (ASF) is a fatal disease caused by a virus in domestic pigs. In this study, a real-time loop-mediated isothermal amplification (LAMP) assay and visual LAMP assay were developed for the detection of African swine fever virus (ASFV). LAMP primers targeting the p10 gene of ASFV were designed, the LAMP reaction system was optimized with plasmid pUC57 containing the p10 gene sequence, and the specificities of the real-time LAMP and the visual assays were tested with the DNA or cDNA of pseudorabies virus (PRV), porcine circovirus type 2 (PCV2), classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine parvovirus (PPV) and ASFV, as well as the plasmid pUC57 containing the p10 gene sequence. The detection limits were determined using a serial dilution of plasmid pUC57 containing the p10 gene sequence. Our results showed that the LAMP assays could accurately and specifically detect ASFV with a detection limit of 30 copies per mu l(-1) of pUC57 containing p10 gene sequence. In addition, the LAMP assays were further evaluated using various genotypes of ASFV strains. Furthermore, the LAMP assays are comparable with the well-established real-time PCR assay. This study provides promising solutions for facilitating preliminary and cost-effective surveillance for prevention and control of ASFV.