犬细小病毒四川株的分离鉴定与一步生长曲线的测定

Isolation and identification of canine parvovirus (CPV) in Sichuan province and determination of one-step growth curve

为研究犬细小病毒（CPV）生长规律，利用F81细胞从四川地区疑似CPV感染的病犬血样粪便中分离得到1株CPV，对该株病毒进行理化试验、微量中和试验、PCR鉴定以及分子生物学系统鉴定后，证实该分离株是CPV，命名为SC—C。以SC—C分离株感染F81细胞，感染后不同时间收取上清，利用CPV荧光定量PCR检测方法对不同样品中病毒DNA进行定量分析，绘制CPV一步生长曲线。结果显示，感染后，0～8h细胞内病毒DNA含量较低，感染8h后，细胞内病毒DNA含量逐渐增长，12～60h细胞内病毒DNA含量迅速增长，60h后细胞内病毒DNA含量变化不大。CPV感染F81细胞，感染后O～12h为潜伏期，病毒DNA含量维持在较低水平；12～60h为突破期，病毒DNA含量呈对数增长；感染60h后增长速度减缓，病毒含量维持在较高水平逐步进入稳定期，病毒DNA含量呈“S型”曲线增长。

To study the growth rhythm of CPV. A canine parvovirus, SC-C strain, was isolated from bloody stool of a sick dog in Sichuan,by cultivation in F81 cells, and the isolated virus was identified as CPV through PCR detection, physical and chemical properties experiment and microneutralization test. The CPV virus was used to infect FS1 cells, and the infected supernatant were collected at different times postinfection. Fluorescent quantitative PCR was used to analyze CPV DNA quantity in different samples, the one-step growth curve of SC-C was determined according to the results. At 0-8 h postinfection, CPV DNA stayed at a low level;after 8 h postinfection,CPV DNA increased slowly in cells,at 12-60 h postin- fection, CPV DNA increased rapidly in cells. While the CPV infected F81 cells, 0-12 h postinfection was a period of incubation,virus DNA stayed at a low level;12-60 h postinfection was a stage of breakthrough, virus DNA employed a logarithmic growth at 60 h postinfection, the growth rate slowed down, virus DNA maintained at a high level and steped into stationary phase gradually. CPV DNA in supernatant was in an s-shaped curve growth.