1株猪传染性胃肠炎病毒的分离鉴定及其生物学特性研究

发布时间：2018-02-12 17:25

  本文关键词： 猪传染性胃肠炎病毒 分离培养 免疫过氧化物酶单层细胞染色法 S基因　出处：《河南农业科学》2017年08期 　论文类型：期刊论文

【摘要】：采集1例疑似感染猪传染性胃肠炎病毒(TGEV)仔猪的粪便,运用RT-PCR方法和血清学方法对病料进行TGEV检测,并应用猪睾丸细胞(ST)进行病毒分离培养,分别采用免疫过氧化物酶单层细胞染色法(IPMA)和RT-PCR方法检测病毒在ST细胞中的增殖动态,应用RT-PCR方法扩增分离TGEV S1基因,经序列测定后,采用DNAStar 7.0和Mega 5.0软件对TGEV进行生物信息学分析。结果表明,病料接种到ST细胞培养后,出现明显细胞病变效应(CPE);RT-PCR检测结果显示,TGEV核酸阳性;血清学鉴定TGEV抗原阳性。将分离毒株命名为TGEV-NY。感染12 h后,IPMA检测结果阳性,且可从细胞培养上清中检出TGEV核酸。序列测定分析表明,分离毒株S1基因开放阅读框(ORF)为2 220 bp,编码740个氨基酸。进化分析显示,分离毒株与我国2006年分离的SC-Y毒株亲缘关系最近,分离毒株属于基因Ⅰ群。
[Abstract]:The feces of a pig suspected to be infected with transmissible gastroenteritis virus (TGEV) were collected. TGEV was detected by RT-PCR method and serological method, and the virus was isolated and cultured by pig testicular cells. The proliferation dynamics of the virus in St cells were detected by immunoperoxidase monolayer staining and RT-PCR method respectively. The S1 gene of TGEV was amplified by RT-PCR method and sequenced. DNAStar 7.0 and Mega 5.0 software were used to analyze TGEV bioinformatics. The results showed that there was a significant cytopathic effect after inoculation with St cells. The results of RT-PCR showed that TGEV nucleic acid was positive. The isolated strain was named TGEV-NY.After 12 hours of infection, TGEV nucleic acid could be detected from the supernatant of cell culture, and the TGEV nucleic acid could be detected from the supernatant of cell culture. The ORF of S1 gene was 2220 BP, encoding 740 amino acids. The phylogenetic analysis showed that the isolated strain was closely related to the SC-Y strain isolated from China in 2006, and the isolated strain belonged to group 鈪，

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