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目的对一株Ⅰ型单纯疱疹病毒(HSV-1)进行分离鉴定和增值特性研究。方法从病人口唇疱液样本中分离得到一株病毒,经噬斑纯化、形态鉴定、序列分析,确定该分离株为HSV-1,被命名为HSVIZW6A。用其分别感染Vero细胞、KMB17细胞、A375细胞,观察其细胞病变情况,并绘制生长曲线,分析病毒的生物学特性。结果 HSVI-ZW6A株病毒以MOI=10感染不同细胞,细胞外和细胞内病毒在0~4h,病毒复制缓慢,4~12h病毒增殖进入对数生长期,12~24h病毒增殖缓慢,进入平台期,但24h之后病毒效价测定显示病毒滴度开始呈现下降的趋势,且细胞外病毒最大量高于细胞内,且HSVI-ZW6A株病毒毒力上高于HSVI-strain17病毒。结论通过本实验成功分离和鉴定了一株Ⅰ型HSV病毒,并对其生物学特性进行了分析,为今后的实验奠定了基础。
Objective To isolate and characterize a strain of herpes simplex virus type 1 (HSV-1). Methods A virus was isolated from a sample of patients’ blister fluid. After plaque purification, morphological identification and sequence analysis, it was identified as HSV-1 and named as HSVIZW6A. The cells were infected with Vero cells, KMB17 cells and A375 cells respectively. The cytopathic effect was observed and the growth curves were drawn. The biological characteristics of the virus were analyzed. Results The virus of HSVI-ZW6A strain infected different cells with MOI = 10. The viruses in the extracellular and intracellular viruses replicated slowly from 0 to 4 hours. The virus proliferated from 4 to 12 hours into the logarithmic growth phase. The virus proliferated slowly from 12 to 24 hours and entered the plateau , However, the titer of virus began to decline after 24h, and the maximum amount of extracellular virus was higher than that of HSVI-ZW6A strain, and the virulence of HSVI-ZW6A strain was higher than that of HSVI-strain17 virus. Conclusions A type Ⅰ HSV virus was successfully isolated and identified by this experiment, and its biological characteristics were analyzed, which laid the foundation for future experiments.