摘要
本研究根据GenBank数据库中登录的非洲马瘟病毒、西尼罗病毒和日本脑炎病毒基因序列(GenBank accession No.:NC_006011,No.:NC_009942和No.:NC_001437),构建重组质粒(命名为pUC57-AHSV,pUC57-WNV和pUC57-JEV)并设计合成特异性引物,采用环介导等温扩增方法(LAMP)进行检测,并进行特异性、敏感性和重复性试验。结果显示,该方法能够在1 h内准确检测靶标,与其他8种核酸均无交叉反应,且最低检测限为10 copies。重复性试验显示,Cq值变异率均小于5%。上述结果表明,本研究建立了一种速度快、成本低、特异性好及灵敏度高的LAMP检测方法,可用于马属动物的有效监测和快速检测。
In this study,recombinant plasmids(named pUC57-AHSV,pUC57-WNV and pUC57-JEV)were con-structed to mimic the viruses according to the gene sequences of GenBank(GenBank accession No.:NC_006011,No.:NC_009942 and No.:NC_001437).Primers were synthesized and performed by loop-mediated isothermal amplification.The specificity,sensitivity,and reproducibility of this detection method were evaluat-ed.The results show that the method can detect the target specifically in one hour and the lowest detec-tion limit reaches 10 copies.The repeatability verification shows that the coefficient of variation of Cq value is less than 5%.In brief,a fast,low-cost,specific,and sensitive LAMP detection method was es-tablished,which can be used for effective monitoring of equines and rapid detection of pathogens.
作者
高家乐
钮冰
陈沁
王艳
GAO Jia-le;NIU Bing;CHEN Qin;WANG Yan(School of Life Sciences,Shanghai University,Shanghai 200444,China;Plant and Food Inspection and Quarantine Technology Center,Shanghai Customs,Shanghai 200135,China)
出处
《中国兽医科学》
CSCD
北大核心
2023年第12期1480-1487,共8页
Chinese Veterinary Science
基金
上海市科技兴农项目(2020-02-08-00-10-F01470)。
关键词
非洲马瘟病毒
西尼罗病毒
日本脑炎病毒
环介导等温扩增
核酸检测
African horse sickness virus
West Nile virus
Japanese encephalitis virus
loop-medi-atedisothermal amplification
nucleicacid test
