犬细小病毒广西流行株的分离鉴定及生物学特性分析

发布时间：2018-04-24 14:35

本文选题：犬细小病毒 + 分离　；参考：《中国兽医科学》2017年07期

【摘要】：为了解当前犬细小病毒广西地方毒株的生物学特性,本研究收集广西各地疑似犬细小病毒(CPV)感染病犬的粪便通过接种F81细胞进行病毒的分离培养,并通过血凝试验、抗性分析、同源性分析和攻毒试验等对分离毒株的生物学特性及致病力进行研究。结果显示,接种3代后,细胞出现明显的病变(CPE),血凝效价为1∶512,第5代病毒的TCID_(50)为1×10~(-3.6)/0.1 m L,能抗热、酸、氯仿和乙醚;电镜观察到CPV为直径约25 nm圆形、无囊膜的结构,应用CPV特异性引物检测到长约1 419 bp和1 254 bp的特异性条带,提示成功分离到一株犬细小病毒毒株;测序分析结果表明,VP2基因与周边国家地区及我国各地方的参考毒株、流行株核苷酸同源性为98.9%~99.8%;系统进化发生分析表明,广西分离株的基因型为CPV-2a亚型,与泰国参考毒株VT148亲缘关系最近;与其它CPV-2a亚型相比,分离株的第267位、305位、324位和440位氨基酸位点发生了变异;动物回归试验结果表明,犬细小病毒广西分离株属于强毒株。本研究成功分离到犬细小病毒广西地方流行株,并对其主要衣壳蛋白VP2基因进行遗传变异分析,为更好的预防和控制犬细小病毒提供依据。
[Abstract]:In order to understand the biological characteristics of local canine parvovirus strains in Guangxi, the feces of dogs infected by suspected canine parvovirus (CPV) were isolated and cultured by inoculation of F81 cells, and the resistance was analyzed by hemagglutination test. The biological characteristics and pathogenicity of isolated strains were studied by homology analysis and virus attack test. The results showed that after 3 passage inoculation, the cells showed obvious pathological changes, the hemagglutination titer was 1: 512, and the TCIDX 50 of the fifth generation virus was 1 脳 10 ~ (-3.6) / 0.1 mL, which could resist heat, acid, chloroform and ether. The structure of CPV was observed to be about 25 nm in diameter and without capsule membrane. The specific bands of about 1 419 BP and 1 254 BP were detected by using CPV specific primers, indicating that a canine parvovirus strain was successfully isolated, and the sequencing analysis showed that VP2 gene was associated with the reference strains in neighboring countries and regions of China. The nucleotide homology of the epidemic strain was 98.9 and 99.8.The phylogenetic analysis showed that the genotype of Guangxi isolate was CPV-2a subtype, which had the closest relationship with Thailand reference strain VT148, and compared with other CPV-2a subtypes. The amino acid loci at the 34th and 440 amino acid loci were mutated at the 267 and 440 position, and the animal regression test showed that the canine parvovirus isolated from Guangxi was highly virulent. In this study, the local canine parvovirus strains in Guangxi were successfully isolated, and the genetic variation of the main capsid protein VP2 gene was analyzed in order to provide a basis for better prevention and control of canine parvovirus.
【作者单位】：广西壮族自治区兽医研究所广西兽医生物技术重点实验室;广西民族大学海洋与生物技术学院;武鸣县动物疫病预防控制中心;
【基金】：国家重点研发计划重点专项(2016YFD0501000)
【分类号】：S852.655

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