中国兽医科学2023，Vol.53Issue(12)：1488-1494,7.DOI:10.16656/j.issn.1673-4696.2023.0212
针对猪瘟病毒Erns基因实时荧光定量PCR检测方法的建立与应用
Establishment and application of TaqMan real-time quantitative PCR for detection of classical swine fever virus Erns gene
摘要
Abstract
In this study,a TaqMan fluorescence quantitative PCR method was developed for the detec-tion of classical swine fever virus(CSFV)Erns gene based on the sequence of CSFV Shimen strain(GenBank ID:AF092448.2).Specificity test,sensitivity test and repeatability test were performed.Clinical samples,CSFV and recombinant viruses were tested.The results showed that the Ct value of TaqMan quantitative PCR assay was in the range of 1 × 109-1 × 102 copies/µL.The correlation coefficient was 0.99,the slope was-3.661 5,and the detection limit was 101 copies/μL.There was no cross reaction with other porcine viruses that caused similar clinical symptoms.The results of repeatability test showed that the coef-ficient of variation between and within groups was less than 2.925 02％,indicating this method has good repeatability.This method is suitable for the identification and detection of clinical samples.This method also applies to CSFV proliferation and recombinant porcine reproductive and respiratory syndrome virus expressing CSFV Erns gene constructed in our laboratory,which is of great significance for the rapid detection of clinical CSFV inf ection,the diagnosis of CSF,the future vaccine research,and the laboratory research on CSFV.关键词
猪瘟病毒/Erns基因/荧光定量PCRKey words
classical swine fever virus(CSFV)/Erns gene/real-time quantitative PCR分类
农业科技引用本文复制引用
王树茂,高飞,王桂花,王安冉,周艳君,姜一峰,乔思娜,赵款,童光志,李丽薇..针对猪瘟病毒Erns基因实时荧光定量PCR检测方法的建立与应用[J].中国兽医科学,2023,53(12):1488-1494,7.基金项目
国家重点研发计划项目(2022YFD1800804,2022YFD1800305) （2022YFD1800804,2022YFD1800305）
国家自然科学基金项目(32072861,31941017) （32072861,31941017）
上海市自然科学基金项目(21ZR1476900,20ZR1469600) （21ZR1476900,20ZR1469600）
