中国畜牧兽医2017，Vol.44Issue(2)：311-318,8.DOI:10.16431/j.cnki.1671-7236.2017.02.002
口蹄疫O型病毒二温式RT-PCR检测方法的建立及初步应用
Development and Application of Two-temperature RT-PCR for Detectionof Type O Foot and Mouth Disease Virus
摘要
Abstract
According to the gene sequences analysis of foot and mouth disease virus (FMDV) in GenBank,a pair of specific primers was designed in the conserved sequence of type O FMDV P1 gene.The reaction parameters were optimized using the uniform design method to develop a two-temperature RT-PCR method for detection of type O FMDV.The results of sensitivity and specificity showed that the two-temperature RT-PCR method was only specific for type O FMDV without amplification of the other viruses.The amplified fragment was same with the expected length.The cloning and sequencing results revealed that the sequence of amplified fragment had 100％ simililarity to the target sequence,and the minimum detection quantity was 1.665 pg/μL,the effective detection rate was consistent with the three step RT-PCR sensitivity test results.54 taper toxicity test pigs were detected,and positive identification results and three-step PCR results was consistent.Compared with the three-step PCR,it could save 20 min.These results indicated that the developed two-temperature RT-PCR for detection of type O FMDV was a kind of accurate,rapid,specific and sensitive detection method.关键词
口蹄疫O型病毒/二温式RT-PCR/检测方法Key words
type O foot and mouth disease virus/two-temperature RT-PCR/detection method分类
农业科技引用本文复制引用
姚怀兵,赵毅,刘宏,张毅,任方,黄炯..口蹄疫O型病毒二温式RT-PCR检测方法的建立及初步应用[J].中国畜牧兽医,2017,44(2):311-318,8.基金项目
自治区科研机构创新发展专项资金(2016D04008) （2016D04008）
自治区产学研联合培养研究生示范基地项目(xjaucxy-yjs-20152008) （xjaucxy-yjs-20152008）
