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

环介导等温扩增 非洲猪瘟病毒 猪细小病毒 猪圆环病毒 病毒学 猪瘟 伪狂犬病 猪繁殖与呼吸综合征病毒 质粒 生物 分子生物学 互补DNA 病毒 基因 遗传学 DNA
作者
Deguo Wang,Jianghan Yu,Yongzhen Wang,Meng Zhang,Li P,Meng Liu,Yanhong Liu
出处
期刊:Journal of Virological Methods [Elsevier BV]
卷期号:276: 113775-113775 被引量:73
标识
DOI:10.1016/j.jviromet.2019.113775
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 μl−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.
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