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目的了解贵州省20例水痘患者流行的水痘-带状疱疹病毒(VZV)主要基因型别,为水痘疫情的防控提供科学依据。方法收集20份临床诊断水痘患者血清标本、咽拭子或水疱液标本,分别用ELISA法检测VZV IgM抗体、实时荧光定量PCR检测VZV特异性核酸、普通PCR扩增VZV开放阅读框(ORF)22核酸片段,用Bio Edit和MEGA 4软件分析测序结果,并与Genbank上VZV参考株序列比较,进行生物信息学分析。结果 20份临床诊断病例VZV IgM抗体检测均为阳性,其中7份咽拭子或水疱液标本实时荧光定量PCR检测为阳性,3例标本的PCR产物在约447bp处有明显目的条带,测序证实为水痘-带状疱疹病毒。ORF22基因的单核苷酸多态性(SNP)分析显示,GZVZV001和GZVZV007核酸片段完全一致,GZVZV002序列在37922位点出现R碱基取代G碱基。3株分离株序列与J基因型参考株p-OKa序列比较,一致性为99%~100%,与E基因型参考株DUMAS序列比较,在6个关键位点上有4个碱基不一致,与M1基因型参考株CA123比较,6个关键位点上有3个碱基不一致。提示3株分离株为VZV J基因型。结论贵州省目前VZV流行的野毒株型别中存在J基因型,加强VZV疫苗的接种能有效防止水痘的爆发流行。
Objective To understand the prevalence of varicella-zoster virus (VZV) in 20 cases of varicella patients in Guizhou province and to provide a scientific basis for the prevention and control of the epidemic of varicella. Methods Serum samples, throat swabs or blister fluid specimens from 20 patients with clinical diagnosis of chickenpox were collected. VZV IgM antibodies were detected by ELISA. VZV specific nucleic acids were detected by real-time fluorescence quantitative PCR. VZV open reading frame (ORF) 22 The nucleic acid fragments were analyzed by Bio Edit and MEGA 4 software and compared with the VZV reference strain on Genbank for bioinformatics analysis. Results Twenty VZV IgM antibodies were detected positive in all of the 20 clinically diagnosed cases. Seven throat swabs or blister fluid samples were positive by real-time fluorescence quantitative PCR, and the PCR products of 3 specimens had obvious target bands at about 447 bp. Sequencing confirmed For chickenpox - herpes zoster virus. Single nucleotide polymorphism (SNP) analysis of the ORF22 gene showed that the GZVZV001 and GZVZV007 nucleic acid fragments were identical, and the G base of the GZVZV002 sequence appeared at the 37922 site. The sequences of the three isolates were 99% ~ 100% identical to the reference strain p-OKa of J genotype. Compared with the DUMAS sequence of E genotype reference strain, there were 4 bases in 6 key sites, Compared with the M1 genotype reference strain CA123, there are 3 bases in 6 key sites. Three strains of VZV J genotypes were suggested. Conclusion There is J genotype in the genotypes of wild VZV in Guizhou Province. Strengthening the vaccination of VZV vaccine can effectively prevent the outbreak of chickenpox.