畜牧兽医学报2018，Vol.49Issue(6)：1314-1319,6.DOI:10.11843/j.issn.0366-6964.2018.06.025
猪圆环病毒3型LAMP检测方法的建立与应用
Establishment and Application of LAMP Method for Detection of Porcine Circovirus Type 3
摘要
Abstract
In order to diagnose and detect porcine circovirus type 3(PCV3),in this research,spe-cific primers were designed according to the conserved sequence of ORF 2 gene.By optimizing the concentration of Bst DNA polymerase,the ratio of primers,Mg2+,dNTPs,betaine and reaction condition,the loop-mediated isothermal nucleic acid amplification(LAMP)was established.The results manifested that ladder strips were observed after 36min with a constant temperature,and the product could be judged by SYBRⅠstaining.The detection limit of this way was 1.0×101copies·μL－1 which had no cross reaction with porcine reproductive and respiratory syndrome virus(PRRSV),porcine pseudorabies virus(PRV),classical swine fever virus(CSFV),porcine circovirus type 1(PCV1)and type 2(PCV2).From 68 lymph nodes of diseased pigs in respiratory system,the positive rate of 30.9％tested by LAM P had a high coincidence rate of 95.6％(65/68)with that by PCR detection.Compared with traditional technology,this method is more sensitive,simpler and can be used for detection and clinical diagnosis of PCV 3.关键词
猪圆环病毒3型/RT-LAMP/检测Key words
PCV3/RT-LAMP/detection分类
农业科技引用本文复制引用
姜辰龙,严秀文,张日腾,李勇,姜平,白娟..猪圆环病毒3型LAMP检测方法的建立与应用[J].畜牧兽医学报,2018,49(6):1314-1319,6.基金项目
国家自然科学基金(31672565 （）
31502082) （）
中央高校基本科研业务费专项资金资助项目(KJQN201616) （KJQN201616）
国家生猪现代产业体系建设专项(CARS-36) （CARS-36）
