广东地区犬瘟热病毒血凝素基因的克隆与序列分析

林希; 刘若寒; 郝香琪; 郑清栩; 陶攀; 周沛; 李守军

doi:10.7671/j.issn.1001-411X.201812044

广东地区犬瘟热病毒血凝素基因的克隆与序列分析

Cloning and sequence analysis of haemagglutinin gene of canine distemper virus in Guangdong area

摘要

摘要:
目的对广州和东莞市宠物犬感染犬瘟热病毒(CDV)的情况进行病原鉴定和分析，为监测CDV的遗传变异情况和防治犬瘟热(CD)提供数据基础。

方法从表现CD症状的犬只中鉴定了17份CDV阳性样本，采用RT-PCR的方法克隆得到这些野毒株的血凝素(H)基因序列，采用生物信息学方法进行序列比对分析。

结果 17株CDV的H基因核苷酸与氨基酸序列的相似性分别为97.4%~100.0%和97.5%~100.0%，与Onderstepoort、Lederle和Convac等疫苗株相比，其核苷酸与氨基酸序列相似性分别为90.3%~91.5%和89.4%~90.8%。进化树分析结果显示，17株CDV野毒株均属于Asia Ⅰ型，与疫苗株的分支较远；本研究鉴定的野毒株已进化形成9个潜在的N−糖基化位点。

结论 Asia Ⅰ型CDV仍为该地区的流行基因型，基因型较稳定，但与疫苗株相比形成了一定的进化距离和出现了大量的变异。因此，继续监控CDV在犬群中的进化，掌握其遗传变异状况具有重要意义。

Abstract:
Objective To perform etiological investigation of canine distemper virus (CDV) infection in dogs from Guangzhou and Dongguan, moniter the genetic evolution of canine CDV, and provide a data basis for the prevention and control of canine distemper (CD).

Method Seventeen CDV positive samples were identified from dogs with CD symptoms. The haemagglutinin ( H ) genes of these wild strains were obtained by RT-PCR, and the H gene sequences were compared and analyzed through bioinformatics approach.

Result The similarities of nucleotide and amino acid sequences of H genes from 17 CDV strains were 97.4% to 100.0% and 97.5% to 100.0%, respectively. Compared with vaccine strains such as Onderstepoort, Lederle and Convac, the similarities of nucleotide and amino acid sequences of H genes from these CDV strains were 90.3% to 91.5% and 89.4% to 90.8%, respectively. Phylogenetic tree analysis showed that 17 CDV wild strains belonged to Asia I subtype and were distant from the vaccine strains. The identified wild strains had evolved to containing nine potential N-glycosylation sites.

Conclusion Asia I CDV is still an epidemic subtype in this region and this subtype is stable, but it has formed a certain evolutionary distance and a large number of mutations compared with vaccine strains. Therefore, it is of great significance to continue to monitor the evolution of CDV in dog populations.

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