期刊论文

Establishment and preliminary application of an indirect ELISA based on S1 protein of porcine epidemic diarrhea virus

中文

中国兽医科学

1673-4696

2020

50

11

1341-1347

10.16656/j.issn.1673-4696.2020.0190

国家自然科学基金面上项目(31672539) 湖南省重点领域研发计划项目(2019NK2171)。

本校为第一机构

根据猪流行性腹泻病毒(PEDV)HuN株S1蛋白的基因序列(GenBank:JQ517274)构建哺乳动物细胞重组表达质粒pIRES-S1。利用电转法将pIRES-S1转染至ST细胞,经过G418加压筛选、Western-blot检测、悬浮驯化后,得到能表达S1蛋白的ST稳转细胞池。Western-blot结果显示,S1蛋白具有良好的反应原性。以S1蛋白作为包被抗原初步建立间接ELISA方法,并对PEDV临床血清和进口种猪PEDV阴性血清进行检测,以血清中和试验为标准,结果显示,该ELISA方法敏感性为96.3%,特异性为97.7%,与血清中和试验具有很高的一致性(kappa值=0.882,P

The recombinant expression plasmid pIRES-S1 was constructed according to the gene sequence of PEDV S1 in NCBI(GenBank:JQ517274).The plasmid pIRES-S1 was transfected into ST cells by electrotransfer.After G418 pressurization screening,western-blot detection and suspension domestication,a stable transduction cell pool expressing S1 protein was obtained.The results of Western-blot showed that S1 protein have good reactivity.An indirect ELISA was established by using S1 protein as coating antigen,and the ELISA was used to detect PEDV clinical serum and PEDV negative serum of imported breeder pigs....