水痘带状疱疹病毒蚀斑减少中和试验

Plaque Neutralization Test of Varicella-Zoster Virus

〔目的〕建立水痘带状疱疹病毒中和抗体测定方法。〔方法〕用VZV1毒种制备中和抗原 ,以 5 0pfu左右病毒量加待测血清 36℃中和 1h ,接种Vero-E6单层细胞 ,烛缸法培养 10d后 ,用结晶紫染色 ,计数pfu ;以中和后pfu减少 5 0 %的血清最高稀释度为中和抗体滴度。〔结果〕VZV1在Vero -E6细胞上形成的蚀斑呈椭圆形 ,平均Φ0 .85mm ,边界清晰 ;经试验 3只VZV1免前家兔血清中和抗体滴度 <1:2 ,免后分别为 1:16、1:31、1:6 4;5份中和抗体阳性血清经重复测试 3次 ,中和抗体最多相差 1个滴度 ;2 4份血清同时测定VZV中和、荧光、酶标抗体的GMT比值为 1:1.83:46 .2 5。〔结论〕VZV蚀斑减少中和试验特异性、重复性良好 ,可用于VZV疫苗免疫效果考核。

Objective〕To establish neutralizing antibodies tset to Varicella-Zoster Virus(VZV). 〔Methods〕 Preparing neutralizing ant igen with VZV1, serial serus dilutions were mixed with an equal volume(100μl) o f diluted VZV1 so as to yield～50pfu per well. After neutralizing for 1hr at 36 ℃, mixture was inoculated to Vero-E6 cells. for 10 days at 36℃. The cells were stained with 0.02% crystal violet in 10% formalin for 5min. The antibody titer w as determined as the dilution of serum that reduced the number of viral plaques per well by 50%.〔Results〕 Vero-E6 cell line had an average plaque size of 0.85mm, Plaques were ellipse and distinct boorder. Before challenge wi th VZV1 neutralizing antibodies, titers of three rabbits were all <1:2, After c ha llenge they were 1;16. 1:32. 1:64, respectively. 〔Conclusions〕 Pla que neutralization test is specific and steady. It can examine immune effect of VZV vaccine.