2021—2022年长春市水痘-带状疱疹病毒流行株基因序列的特征分析

Genetic sequence characteristics of varicella-zoster virus epidemic strains in Changchun City from 2021 to 2022

目的对2021—2022年长春市水痘-带状疱疹病毒(varicella-zoster virus,VZV)流行株进行基因型鉴定及分子特征分析。方法选择59例2021—2022年吉林大学中日联谊医院收治的长春市水痘或带状疱疹患者为研究对象,采用膜抗原荧光抗体(fluorescent antibody to membrane antigen,FAMA)法对患者血清中VZV特异性抗体进行鉴定。提取确诊患者疱疹液中的病毒DNA,PCR扩增VZV的多个开放阅读框(open reading frame,ORF),包括ORF1、ORF12、ORF16、ORF17、ORF21、ORF22、ORF37和ORF54,根据ORF的单核苷酸多态性(single nucleotide polymorphism,SNP)进行基因分型。通过限制性片段长度多态性(restriction fragment length polymorphism,RFLP)技术对多个ORF(ORF38、ORF54和ORF62)进行酶切鉴定,区分VZV临床株及疫苗株。结果59份患者血清样本均为阳性,均确诊为VZV感染。59份疱疹液样本中,4份与Clade 2基因型完全匹配;4份存在2个SNPs,分别为ORF1(SNP790)C→A突变和ORF12(SNP18082)T→C突变;51份发生ORF12(SNP18082)T→C突变。59株VZV临床株均为PstⅠ^(+)BglⅠ^(+)SmaⅠ^(-),不同于疫苗株PstⅠ^(-)BglⅠ^(+)SmaⅠ^(+)。结论2021—2022年长春市VZV流行株均为Clade2基因型,未发现疫苗株致病情况。

Objective To identify the genotypes and analyze the molecular characteristics of varicella-zoster virus(VZV)endemic strains in Changchun from 2021 to 2022.Methods A total of 59 patients with varicella or herpes zoster treated in China Japan Union Hospital of Jilin University in Changchun from 2021 to 2022 were selected as the research objects,which were identified for the serum VZV-specific antibodies by fluorescent antibody to membrane antigen(FAMA)method.The viral DNA was extracted from herpetic fluid of the confirmed patients,and multiple open reading frames(ORFs)including ORF1,ORF12,ORF16,ORF17,ORF21,ORF22,ORF37,and ORF54 of VZV were amplified by PCR,which was genotyped according to the single nucleotide polymorphisms(SNPs)of the ORFs.Furthermore,multiple ORFs,ORF38,ORF54 and ORF62 were identified by using restriction fragment length polymorphism(RFLP)to distinguish clinical strains and vaccine strains.Results 59 serum samples were positive,indicating that all the patients were infected with VZV.Among the 59 clinical samples,4 samples were completely matched with Clade 2 genotype,4 samples showed 2 SNPs,including ORF1(SNP790)mutation of C→A and ORF12(SNP18082)mutation of T→C,and 51 samples showed ORF12(SNP18082)mutation of T→C.All 59 VZV clinical isolates were PstⅠ^(+)BglⅠ^(+)SmaⅠ-,which was different from the vaccine strain PstⅠ-BglⅠ^(+)SmaⅠ^(+).Conclusion All the VZV endemic strains in Changchun from 2021 to 2022 were Clade 2 genotype,and no vaccine strain was found to be pathogenic.