水痘带状疱疹病毒培养液的筛选

Screening of medium for varicella-zoster virus

目的筛选培养水痘带状疱疹病毒(varicella-zoster virus,VZV)的培养液。方法将4种不同的培养基分别与新生牛血清、人血白蛋白组合,配制12组不同的病毒培养液,培养VZV后,采用噬斑法检测病毒滴度。用筛选出的病毒培养液,对VZV进行6个40层细胞工厂的扩大化培养及连续传代培养,收获P1和P5病毒,进行ORF62核酸序列测序,利用DNAMan软件对测序结果进行比对分析。结果实验2、4、11组培养液培养的VZV滴度明显高于其他组(P均<0.05),选择实验4组培养液对VZV进行扩大化培养后,病毒滴度均稳定在约5.4 Lg PFU/ml;连续传代培养,ORF62核酸序列无碱基突变。结论筛选出1种不含血清及人血白蛋白的VZV培养液,可以稳定地培养VZV。

Objective To screen the culture medium for varicella-zoster virus（VZV）. Methods Twelve groups of virus culture media were formulated with four kinds of media and newborn bovine serum or human serum albumin, with which VZV was cultured and determined for titer. By using the screened medium, VZV was subjected to expand culture and continuous subculture in six 40-layer cell factory. The VZV of P1 and P5 were harvested, of which the ORF62 was sequenced, and the result was analyzed by using DNAMan software. Results The titers of VZV in media of groups 2, 4 and 11 were significantly higher than those in other groups（each P 〈 0. 05）. The medium of group 4 was screened for expand culture, in which the VZV titer was stabilized at about 5. 4 Lg PFU/ml. No base mutation was found in the nucleic acid sequence of ORF62 of VZV after continuous subculture. Conclusion A serum-and human serum albumin-free medium was screened, which was suitable for stable culture of VZV.