猪流行性腹泻病毒双重RT-PCR检测方法的建立
Development of a Duplex RT-PCR Targeting Both N Gene and S Gene for Detection of Porcine Epidemic Diarrhea Virus
DOI: 10.12677/AMB.2017.64017, PDF,    科研立项经费支持
作者: 桑益旸, 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
关键词: 猪流行性腹泻病毒；双重RT-PCR；N基因；S基因；Porcine Epidemic Diarrhea Virus； Duplex RT-PCR； N Gene； S Gene
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(Porcine Epidemic Diarrhea Virus, PEDV)的双重RT-PCR检测方法。在20 μL反应体系中，该双重RT-PCR可检测105拷贝的PEDV cDNA，检测传染性胃肠炎病毒、繁殖与呼吸障碍综合征病毒、伪狂犬病毒和圆环病毒2型、乙型脑炎病毒、猪瘟病毒等几种猪病疫苗毒的结果均为阴性，表明该方法具良好的特异性和灵敏性。临床样品检测结果表明该双重RT-PCR能满足临床检测需要，可一定程度弥补单重RT-PCR漏检的不足。
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 encephalitis B (SEEBV), Swine fever virus (SFV), showing high sensitivity and specificity. The results of clinical sample detection indicated that the established duplex RT-PCR could meet clinical detection and make up the defect of singleplex RT-PCR.
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. https://doi.org/10.12677/AMB.2017.64017

参考文献

[1] Pensaert, M.B., Debouck, P. and Reynolds, D.J. (1981) An Immunoelectron Microscopic and Immunofluorescent Study on the Antigenic Relationship between the Coronavirus-Like Agent, CV 777, and Several Coronaviruses. Archives of Virology, 68, 45-52. [Google Scholar] [CrossRef] 
[2] Sun, R.Q., Cai, R.J., Chen, Y.Q., et al. (2012) Outbreak of Porcine Epidemic Diarrhea in Suckling Piglets, China. Emerging Infectious Diseases, 18, 161-163. [Google Scholar] [CrossRef] [PubMed]
[3] Sun, M., Ma, J.L., Wang, Y.N., et al. (2015) Genomic and Epidemiological Characteristics Provide New Insights into the Phylogeographical and Spatiotemporal Spread of Porcine Epidemic Diarrhea Virus in Asia. Journal of Clinical Microbiology, 53, 1484-1491. [Google Scholar] [CrossRef] 
[4] Stevenson, G.W., Hoang, H., Schwartz, K.J., et al. (2013) Emergence of Porcine Epidemic Diarrhea Virus in the United States: Clinical Signs, Lesions, and Viral Genomic Sequences. Journal of Veterinary Diagnostic Investigation, 25, 649-654.
[5] Kocherhans, R., Bridgen, A., Ackermann, M., et al. (2001) Completion of the Porcine Epidemic Diarrhoea Coronavirus (PEDV) Genome Sequence. Virus Genes, 23, 137-144. [Google Scholar] [CrossRef] 
[6] Lin, C.-M., Saif, L.J., Marthaler, D. and Wang, Q.H. (2016) Evolution, Antigenicity and Pathogenicity of Global Porcine Epidemic Diarrhea Virus Strains. Virus Research, 226, 20-39. [Google Scholar] [CrossRef] [PubMed]

为你推荐