广西黄牛乳头状瘤病毒的鉴定及其基因组序列分析

Identification of bovine papillomavirus and genome sequence analysis for cattle in Guangxi

  • 摘要: 目的明确广西黄牛是否存在牛乳头状瘤病毒(Bovine papillomavirus,BPV)感染,为今后制定科学的防控措施提供参考依据.方法从广西隆安县某黄牛养殖场青年黄牛的皮肤上采集瘤状物病料,在临床观察和组织病理学观察的基础上,采用PCR对其进行鉴定和基因型分型,再根据鉴型结果设计5对特异性引物对BPV基因组进行扩增及测序分析.结果黄牛皮肤瘤状物的病理组织切片观察发现,表皮与真皮增生,表皮细胞角质化和空泡化,细胞呈梭形或星形.从瘤状物中分离获得1株毒株,其与BPV-1型的同源性达99.0%,命名为BPV-GX-LA150909.BPV-GX-LA150909毒株基因组全长7946 bp,GenBank登录号为MF435918,包含早期基因区(E区)、晚期基因区(L区)和非编码区(NCR区)3个区域.BPV-GX-LA150909毒株无论是其基因组还是L1基因,均与BPV-1型的同源性最高,尤其与贵州株BPV-1 GZLZ的同源性分别为99.9%和99.7%,且在系统发育进化树上也是与BPV-1 GZLZ的遗传距离较近.BPV-GX-LA150909毒株的L1蛋白具有潜在的B细胞抗原表位,含有17个O-糖基化位点和1个N-糖基化位点.结论广西黄牛群中已存在BPV-1型感染,且与国内外BPV-1型的同源性较高,说明BPV跨地区传播是不可忽视的问题.

     

    Abstract: ObjetiveWhether cattle in Guangxi have been infected by bovine papillomavirus(BPV)was investigated to provide reference for formulating prevention and control measures of BPV in the future.MethodTumor-shaped sam-ples were collected from the skin of young cattle in a cattle farm in Longan,Guangxi.Based on clinical observation and histopathological observation,BPV were identified and their genotypes were classified by PCR.Then five pairs of speci-fic primers were designed to amplify and sequence BPV genome according to the previous results.ResultHistopathologi-cal slices of the tumor-shaped tissues revealed that,it had epidermis and dermis hyperplasia,and epidermal cells keratini-zation and cavitation occurred.The cells were fusiform and stellate.One strain was isolated from the tumor-shaped sam-ple. Its homology with BPV-1 was 99.0%,and named BPV-GX-LA150909.The full-length genome of strain BPV-GX-LA150909 was 7946 bp,and its GenBank accession number was MF435918.It included three regions:early gene region (E),late gene region(L)and non-coding region(NCR).The homology between strain BPV-GX-LA150909 and BPV-1 type in both genome and L1 gene was the highest,especially with Guizhou strain BPV-1 GZLZ(genome homology 99.9%,L1 gene homology 99.7%). In addition,phylogenetic tree showed that genetic distance of this strain was also close to BPV-1 GZLZ.L1 protein of BPV-GX-LA150909 presented potential B cell epitope,and contained seventeen O-glycosylation sites and one N-glycosylation site.ConclusionBPV-1 infection has been found in Guangxi cattle,and the homology with BPV-1 type at home and abroad is high,which indicates that cross-regional transmission of BPV is a prob-lem that cannot be ignored.

     

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