猪流行性腹泻病毒双重RT—PCR检测方法-建立.pdf

Advances in Microbiology 微生物前沿, 2017, 6(4), 131-136 Published Online December 2017 in Hans. /journal/amb /10.12677/amb.2017.64017 Development of a Duplex RT-PCR Targeting Both N Gene and S Gene for Detection of Porcine Epidemic Diarrhea Virus Yiyang Sang, Hao Wu, Guanglan Tu, Tao Sun, Heng’an Wang* Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiaotong University, Shanghai th th th Received: Dec. 6 , 2017; accepted: Dec. 20 , 2017; published: Dec. 27 , 2017 Abstract To improve the correct probabilities for detection of clinical samples, a duplex RT-PCR targeting both N gene and S gene for detection of Porcine Epidemic Diarrhea Virus (PEDV) was developed. In a 20 μL reaction system, 105 copies of PEDV cDNA could be detected and with no cross reaction with Transmissible Gastroenteritis Virus (TGV), Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), Pseudorabies Virus (PRV), Porcine Circovirus 2 (PCV2), Swine epidemic encephali- tis B (SEEBV), Swine fever virus (SFV), showing high sensitivity and specificity. The results of clin- ical sample detection indicated that the established duplex RT-PCR could meet clinical detection and make up the defect of singleplex RT-PCR. Keywords Porcine Epidemic Diarrhea Virus, Duplex RT-PCR, N Gene, S Gene 猪流行性腹泻病毒双重RT-PCR检测方法的 建立 旸 * 桑益 ，吴 昊，涂光岚，孙 涛，王恒安 上海交通大学农业与生物学院，上海市兽医生物技术重点实验室，上海 收稿日期：2017年12月6 日；录用日期：2017年12月20 日；发布日期：2017年12月27 日 *通讯作者。 文章引用: 桑益旸, 吴昊, 涂光岚, 孙涛, 王恒安. 猪流行性腹泻病毒双重 RT-PCR 检测方法的建立[J]. 微生物前沿, 2017, 6(4): 131-136. DOI: 10.12677/amb.2017.64017 桑益旸 等 摘 要 为提高临床检测的准确性，基于N基因和S基因，建立了检测猪流行性腹泻病毒(Porci