中国兽医科学2012，Vol.42Issue(6)：601-605,5.
猪瘟病毒NASBA-ELISA检测方法的建立与应用
Establishment and application of an NASBA-ELISA for detection of classical swine fever virus
摘要
Abstract
In order to establish a nucleic acid sequence-based amplification（NASBA） method for the detection of classical swine fever virus（CSFV）,specific NASBA primers and probe of CSFV E2 gene were designed using Primer 5.0 and Blastn. Based on solution hybridization and enzyme-linked technology, the NASBA-ELISA method for the detection of CSFV were developed successfully,and the optimized amplification condition of nucleic acid isolation is 42 ℃ and 1.5 h. The result of clinical detection indicated that specific fragments of 155 bp were obtained from the CSFV positive samples by NASBA, other than in TGEV,PRRSV,PCV2,PPV,PEDV,APP and PK-15 cell samples. The NASBA assay had a detection limit equivalent from 1 to 10 CSFV copies/μL. The sensibility of NASBA-ELISA was similar to that of the RT-PCR. A total of 125 clinical samples of pigs were analyzed for CSFV with a positive rate of 5.6 % ,which showed that this assay had a coincidence rate of 94. 4% with NASBA-ELISA and RT-PCR. In conclusion, this NASBA-ELISA method was simple to perform and can be applied to the rapid identification and detection of CSFV.关键词
猪瘟病毒/E2基因/NASBA-ELISA方法Key words
classical swine fever virus/E2 gene nucleic acid sequence-based amplification diagnosis method-ELISA分类
农业科技引用本文复制引用
聂福平,肖进文,李应国,徐自忠,王灵强,王昱,周雪梅,李小兰..猪瘟病毒NASBA-ELISA检测方法的建立与应用[J].中国兽医科学,2012,42(6):601-605,5.基金项目
国家质检总局科研项目 （）
重庆市自然科学基金项目 （）
