中国兽医科学2017，Vol.47Issue(5)：544-550,7.DOI:10.16656/j.issn.1673-4696.2017.05.002
猪繁殖与呼吸综合征病毒与猪流行性腹泻病毒双重RT-PCR检测方法的建立与应用
Establisment and application of a double RT-PCR for the detection of PRRSV and PEDV
摘要
Abstract
In order to detect PRRSV and PEDV rapidly,we designed a pair of specific primers according to the published gene sequences of PRRSV ORF7 gene and PEDV S gene in GenBank.By optimizing the components of the reaction system,a double RT-PCR method was developed for simultaneous detection of PRRSV and PEDV.The method could amplify specific fragments of 300 bp and 578 bp from the mixed genome of PRRSV and PEDV,respectively and the results of TGEV,CSFV,JEV,RVand E.coli were negative.This ruslts showed that the specificity was good.We detected 79 clinical samples,and the results were consistent with single RT-PCR,so this method had good repeatability.The detection limit of this method was 1.49 × 105 copies/μL,which has good sensitivity.The establishment of this method for PRRSV and PEDV differential diagnosis and rapid detection of mixed infections has an important value.The duplex RT-PCR method established in this study has good practicability and can be used for the detection of clinical samples.关键词
猪繁殖与呼吸综合征病毒/猪流行性腹泻病毒/ORF7基因/S基因/双重RT-PCRKey words
PRRSV/PEDV/0RF7 gene/S gene/double RT-PCR分类
农业科技引用本文复制引用
李小璟,龚双燕,陈瑛琪,李雨濛,蔡瑶,徐逸飞,朱玲,徐志文..猪繁殖与呼吸综合征病毒与猪流行性腹泻病毒双重RT-PCR检测方法的建立与应用[J].中国兽医科学,2017,47(5):544-550,7.基金项目
四川省科技支撑计划项目(2017NZ0038) （2017NZ0038）
“十二五”农村领域国家科技计划项目(2015BAD12B04-2.3) （2015BAD12B04-2.3）
