2005-2010年北京地区儿童原发性EBV感染流行株EBNA1与LMP1基因特征分析

摘要

Objective To analyze the characteristic of nuclear antigen 1 gene and latent membrane protein 1 gene of Epstein-Barr virus in primary EBV infection in children in Beijing area in 2005-2012.Methods Polymerase chain reaction (PCR) was used to amplify the EBNA-3C,EBNA1 and LMP1 genes.The amplified products were sequenced directly and the sequences were analyzed by BioEdit 7.0.9 and MEGA 4.0.2.Results Type A EBV was detected in 98％ samples.Nucleotide sequence analysis of the carboxy-terminal region of EBNA1 showed that Vvv1 was deteted in 98％ samples.DNA sequence analysis of LMP1 C-terminus indicated that China 1 was 90％ in this study.There were no significant differences in the frequency of Vvv1 and China 1 between the IM and HLH samples (P =1.00).Linkage analysis of EBV types,EBNA1 and LMP1 variants indicated that 90％ of EBV type A was associated with EBNA1-Vvv1 variant and LMP1-China 1 variant in 40 cases.Full length of LMP1 gene was successfully amplified in 35 cases.Four Chinese groups (CG1-4) were identified.The percentage of CG1-CG4 were 85％,6％,6％ and 3％,respectively.Conclusion EBV type A is predominant in primary EBV infection in children in Beijing Area.EBNA1-Vvv1 and LMP1-China 1 variants were predominant genotypes in this area.There is a high linkage between EBNA1-Vvv1 variant and LMP1-China 1 variant.Four Chinese groups (CG1-4) were identified according to the full length of LMP1 gene and CG1 was the most prevalent.%目的 分析2005-2010年北京地区儿童原发性EBV感染流行株EBNA1与LMP1基因特征,为研究EBV变异株与疾病临床表型间是否存在相关性提供背景资料.方法 应用PCR方法扩增EBV的EBNA3C、EBNA1和LMP1基因片段,测序后应用BioEdit 7.0.9和Mega 4.0.2软件进行序列分析.结果 62例进行了EBV分型,以EBV-Ⅰ型为主,检出率为98％.62例EBNA1基因扩增阳性,其中V-val亚型(均是Vvv1变异株)为98％.50例LMP1基因羧基段扩增阳性,以China 1为主,其检出率为90％.Vvv1变异株和China 1变异株在EBV-IM与EBV-HLH中的检出率差异均无统计学意义(P=1.00).40例进行了多基因连锁分析,其中EBV-Ⅰ型、EBNA1-Vvv1变异株和LMP1-China 1变异株高度连锁,其连锁检出率为90％.35例患儿LMP1基因全长扩增阳性,CG1-CG4的检出率分别为85％、6％、6％和3％.结论 北京地区儿童原发性EBV感染疾病中,EBV亚型以EBV-Ⅰ型为主,Vvv1变异株和China 1变异株分别是EBNA1和LMP1变异株中的优势变异株,且二者高度连锁.儿童原发性EBV感染流行株可以分为CG1-4组,其中以CG1为主.