摘要
为了解目前流行的猪传染性胃肠炎病毒（TG EV）的生物学特征及其遗传变异情况,从疑似感染TGEV的猪群中采集发病仔猪小肠,病料处理后在ST细胞上盲传,至第5代出现特征性细胞病变,通过PCR和间接免疫荧光验证,成功分离出2株TGEV,被命名为H E-1和HE-4。RT-PCR方法扩增测序其S基因,对14株TGEV S基因进化和变异情况进行分析。氨基酸比对发现,与TGEV经典毒株Purdue株相比,分离株在第562~564 aa处缺少N NT糖基化位点,与TS株相比在第375~377 aa处缺少ND T糖基化位点;同时,HE-4株在影响抗原位点D位点的第385 aa处发生突变。这种糖基化位点的缺失和抗原位点的突变,对TGEV生物学特性的影响有待进一步研究。
In order to further research the characteristics of the current transmissible gastroenteritis virus of swine（TGEV） and its different genetic mutations from the traditional strains, collections of specimens from the suspected pigs with TGEV infection were cultured in the ST cells. After five passages, the typical cytopathic effect was observed. By RT-PCR and indirect immunofluorescence test, two TGEV isolates were identified successfully and named as HE-1 and HE-4, respectively.Then,to analyse the evolution and variation of 14 strains the S genes were amplified and sequenced by RT-PCR The results showed that during the alignment of nucleotide and amino acid, their 562—564aa NNT glycosylation site was deleted compared with the classical TGEV Purdue strain; and their 375-377 aa NDT glycosylation site was deleted compared with the TS strain. In addition, it was found that HE-4 strain mutated at 385 aa on the major antigen site D. Such deletions on the glycosylation sites and the mutation on the major antigen sites are required to the further research into the biological characteristics of TGEV.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2016年第3期296-302,共7页
Chinese Veterinary Science
基金
国家自然科学基金项目(31372438
31200122)
教育博士点基金项目(20122325110019)
哈尔滨市科技局"优秀学科带头人"研究专项资金项目(2012RFXXN009)
关键词
猪传染性胃肠炎病毒
分离鉴定
S基因
分子生物学特征
transmissible gastroenteritis virus of swine
isolation and identification
S gene
molecular biological characteristics
