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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 for the culture of varicella-zoster virus (VZV). Methods Four kinds of different media were respectively combined with newborn calf serum and human serum albumin to prepare 12 groups of different virus culture solutions. After culturing VZV, the virus titer was detected by plaque method. VZV was expanded and cultured continuously in six 40-layer cell factories with the selected virus culture solution. P1 and P5 viruses were harvested and ORF62 nucleic acid sequence was sequenced. DNAMan software was used to compare the sequencing results. Results The titer of VZV in experiment 2,4,11 group was significantly higher than that in other groups (P <0.05). After the experiment group 4 culture medium was expanded to VZV culture, the titer of VZV was stable at about 5.4 Lg PFU / ml; continuous subculture, ORF62 nucleic acid sequence without base mutation. Conclusion A VZV culture medium without serum and human serum albumin was screened out to stabilize VZV.