儿童咽拭子标本406份中EB病毒的检测及其基因型鉴定

Detection and genotyping identification of Epstein-Barr virus nasopharyngeal swabs in children

目的了解湖南省株洲、湘潭和衡阳地区儿童鼻咽部EB病毒的感染状况及其基因型，为EB病毒的治疗、预防和临床检验提供指导。方法从株洲、湘潭和衡阳部分医院收集疑似EB病毒感染的儿童咽拭子标本406份，采用荧光定量聚合酶联反应（FQ—PCR）检测标本中的EB病毒-DNA；阳性标本分别用特异性引物从中扩增3个目的基因片段，其中PCR扩增的EBNA-3C片段直接电泳以分析1／2分型，BamHI片段和BamHI WI／I1交界区基因片段采用PCR-RFLP分析鉴定F／f和C／D分型，测序和Blast比对验汪分型结果。结果从406份标本中共检出 EB病毒-DNA阳性株159份，株洲74份（38．5％），其中1型72份（97．3％），2型2份（2．7％）；湘潭42份（38．9％），1型40份（95．2％），2型2份（4．8％）；衡阳43份（40．6％），1型43份（100％），2型未发现、从159份阳性株中随机扩增了73份做F／f分型，68份做C／D分型，共检测到F型73份，C型68份，3个地区均未检测到f型和D型。结论株洲，湘潭和衡阳儿童鼻咽部EB病毒感染的检出率分别为38．5％，38．9％，40．6％，且其主要感染的3种独立基因型以1、C、F型为主，3个地区间儿童鼻咽部EB病毒-DNA的检出率及其基因型的差异无统计学意义。

Objective To know the state and genotyping of Epstein-Barr virus（EBV） in^ection of naso- pharyngeal swabs in children. Methods Totally 406 samples were detected for EBV-DNA by RQ-PCR. The samples were collected fi＇om nasopharyngeal swabs of children passibly suffering from Epstein-Ban＂ Virus infection in Zhuzhou, Xiangtan and Hengyang cities in Hunan province in China. The analysis of genotypes was imp｜emented on three regions of EBV genome; EBNA-3C, BamHl WI/II and BamHI Fwere detected by the polymerase chain reaction （PCR） and restriction fragment length polymorphism （ RFLP）. Results Totally 159 clinical strains were i- dentified fi＇om children nasopharyngeal swabs. The EBV positive strains were detected in 74 of 192, 42 of 108,43 of 106 samples from Zhuzhou, Xiangtan and Hengyang, respectively. The genotype I was proven in 74/192, 42/ 108, 43/106 clinical samples in those areas, respectively. The frequencies of genotype F and genotype C were de- tected in all cases （ 100% ）. Conclusions The frequencies of EBV-DNA are 38.5% , 38.9% and 40.6% ; genotypes l , C and F are the predonfinant genotyping of EBV. The frequencies of EBV-DNA and the type 1/2, type C/D, type F/f are not significantly different.