广州地区猫泛白细胞减少症病毒分离鉴定及全基因组遗传分析

Isolation,identification and sequence analysis of complete genome of feline panleukopenia virus in Guangzhou area

【目的】分析广州地区猫泛白细胞减少症病毒(Feline panleukopenia virus,FPLV)的自然重组、跨宿主传播以及流行变异情况。【方法】采集疑似感染猫泛白细胞减少症病毒的猫粪便样品进行细胞分离,并对阳性病料提取基因组DNA,PCR扩增获得病毒的全基因组序列,并与Gen Bank中相关的参考毒株序列进行遗传进化分析,同时对VP2基因与NS1基因的主要氨基酸位点进行差异分析。【结果】成功获得2株猫泛白细胞减少症病毒及其全基因组序列。遗传进化分析显示,广州地区分离的猫泛白细胞减少症病毒FPLV-GZ01、FPLV-GZ02与我国其他分离毒株FPLV-XJ01、FPLV-HRB属同一分支,提示FPLV-GZ01、FPLV-GZ02由FPLV-XJ1进化而来;NS1基因系统进化树显示,FPLV存在与CPV-447重组的可能。主要氨基酸位点分析显示,FPLV在VP2基因中的主要氨基酸位点上的遗传变异比CPV保守;在NS1基因上发现FPLV-GZ01、FPLV-GZ02分别存在不同程度的氨基酸位点突变。【结论】广州地区分离的猫泛白细胞减少症病毒仍在不断地重组进化。

【Objective 】 To study the natural recombination,cross host transmission and prevalence of feline panleukopenia virus（FPLV） in Guangzhou area.【Method】Cells were isolated from stool samples of cats seemingly infected with FPLV,genome DNAs were extracted from positive samples,and complete virus genome sequences were amplified by PCR and were compared with relevant reference sequences from Gen Bank using genetic evolution analysis.The major amino acid sites were analyzed for differences between VP2 gene and NS1 gene.【Result】Two FPLV strains were successfully isolated and the complete genome sequences of FPLV were obtained.The genetic evolution analysis indicated that FPLV-GZ01 and FPLV-GZ02 which were isolated from Guangzhou area belonged to the same branch with the other isolated strains FPLV-XJ1 and FPLV-HRB,and implied FPLV-GZ01 and FPLV-GZ02 evolved from FPLV-XJ01.The NS1 gene phylogenetic analysis demonstrated that FPLV might have recombined with CPV-447.Analysis of major amino acid sites indicated that FPLV was more conservative than CPV in the genetic variation of the VP2 gene.FPLV-GZ01 and FPLV-GZ02 had different degrees of mutation in the amino acid sites of the NS1 gene.【Conclusion】The feline panleukopenia viruses isolated from Guangzhou area have been still recombining and evolving.