本文關(guān)鍵詞： 豬傳染性胃腸炎病毒 分離培養(yǎng) 免疫過(guò)氧化物酶單層細(xì)胞染色法 S基因　出處：《河南農(nóng)業(yè)科學(xué)》2017年08期 　論文類型：期刊論文

【摘要】：采集1例疑似感染豬傳染性胃腸炎病毒(TGEV)仔豬的糞便,運(yùn)用RT-PCR方法和血清學(xué)方法對(duì)病料進(jìn)行TGEV檢測(cè),并應(yīng)用豬睪丸細(xì)胞(ST)進(jìn)行病毒分離培養(yǎng),分別采用免疫過(guò)氧化物酶單層細(xì)胞染色法(IPMA)和RT-PCR方法檢測(cè)病毒在ST細(xì)胞中的增殖動(dòng)態(tài),應(yīng)用RT-PCR方法擴(kuò)增分離TGEV S1基因,經(jīng)序列測(cè)定后,采用DNAStar 7.0和Mega 5.0軟件對(duì)TGEV進(jìn)行生物信息學(xué)分析。結(jié)果表明,病料接種到ST細(xì)胞培養(yǎng)后,出現(xiàn)明顯細(xì)胞病變效應(yīng)(CPE);RT-PCR檢測(cè)結(jié)果顯示,TGEV核酸陽(yáng)性;血清學(xué)鑒定TGEV抗原陽(yáng)性。將分離毒株命名為T(mén)GEV-NY。感染12 h后,IPMA檢測(cè)結(jié)果陽(yáng)性,且可從細(xì)胞培養(yǎng)上清中檢出TGEV核酸。序列測(cè)定分析表明,分離毒株S1基因開(kāi)放閱讀框(ORF)為2 220 bp,編碼740個(gè)氨基酸。進(jìn)化分析顯示,分離毒株與我國(guó)2006年分離的SC-Y毒株親緣關(guān)系最近,分離毒株屬于基因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，

本文編號(hào)：1506166