福建农林大学学报（自然科学版）2018，Vol.47Issue(2)：212-216,5.DOI:10.13323/j.cnki.j.fafu(nat.sci.).2018.02.013
猪轮状病毒TaqMan实时荧光定量RT-PCR方法的建立
Establishment of a TaqMan based real-time RT-PCR method for porcine rotavirus
摘要
Abstract
In order to develop a TaqMan based real-time RT-PCR method for porcine rotavirus,specific primers and probe were de-signed by Oligo7 software targeting the sequences of VP6 gene for porcine rotavirus. The established real-time RT-PCR method could detect at 192 copies.μL-1with high sensitivity. No positive signal was detected from common porcine infectious diseases such as porcine epidemic diarrhea virus,transmissible gastroenteritis virus of swine,classical swine fever virus and Japanese encephalitis vi-rus. A standard curve generated had a wide dynamic range with a linear correlation(R2) of 0.999. Excellent reproducibility was ob-tained for detecting constructed positive plasmids with intra-assay of 0.25%-1.26% and inter-assay of 0.31%-1.49%,respectively. Seventy-nine clinical samples were tested by the development TaqMan based real-time RT-PCR method and conventional RT-PCR method with the positive ratio were 5.06%(4/79) and 7.59%(6/79),respectively. RT-PCR method positive samples were all test-ed positive by the TaqMan based real-time RT-PCR method,with coincidence rate was 100%.The TaqMan based real-time RT-PCR method established in this study may be used for earlier diagnosis for porcine rotavirus.关键词
猪轮状病毒/TaqMan/实时荧光定量RT-PCR方法Key words
porcine rotavirus/TaqMan/real-time RT-PCR method分类
农业科技引用本文复制引用
陈如敬,章秋月,陈秋勇,修金生,吴学敏,严山,周伦江..猪轮状病毒TaqMan实时荧光定量RT-PCR方法的建立[J].福建农林大学学报（自然科学版）,2018,47(2):212-216,5.基金项目
福建省生猪产业体系项目(2017No.129) （2017No.129）
福建省属公益类科研专项(2018R1023-92015R1102) （2018R1023-92015R1102）
福建省农业科学院科技下乡"双百"行动项目(sbmb1701) （sbmb1701）
福建省农科院重点创新团队(STIT2017-1-5). （STIT2017-1-5）
