柯萨奇病毒B组4型河南分离株全基因组序列分析

Genomic characteristics of coxsackievirus B4 isolated in Henan province of China

目的了解河南省手足口病病原柯萨奇病毒B组4型(coxsackievirus B4,CV-B4)全基因组特征,分析不同疾病来源CV-B4病毒分离株VP1区基因异同。方法对采集的手足口病患者临床标本134份进行RT-PCR扩增和病毒分离鉴定;对其中1株分离株通过4对引物分段扩增,拼接CV-B4分离株基因组序列,用生物信息学软件分析,构建序列遗传树。结果测序获得河南省CV-B4分离株HN23/CB4/CHN/2013基因组全长序列7 394 bp,5'非编码区(5'-UTR)、P1、P2、P3、3'非编码区(3'-UTR)区域核苷酸序列与Gen Bank公布的其他分离株相似性分别为80.3%~95.7%、80.8%~93.9%、78.6%~87.7%、78.1%~81.2%和66.7%~87.5%;VP1区核苷酸相似性为81.2%~93.2%,氨基酸相似性为96.1%~99.3%,与糖尿病患者分离株E2核苷酸差异最大,达28.8%,与手足口病分离株差异最小,为6.8%。结论 CV-B4病毒同种疾病分离株VP1区域核苷酸同源性较高,不同疾病分离株差异性较大。

Objective To study the genomic characteristics of coxsakievirus B4（CV-B4） strain isolated from patients with hand,foot and mouth disease（HFMD） in Henan province of China. Methods A total of 134 samples were detected by reverse-transcription polymerase chain reaction（RT-PCR） and CV-B4 were isolated by cell culture. The whole genome of one CV-B4 strain was amplified using 4 pairs of primers,the sequences were analyzed and phylogenetic tree was generated by bioinformatics software. Results The full length of HN23/CB4/CHN/2013 genome was 7 394 bp. Compared with the other CV-B4 strains released in Gen Bank,the similarities of nucleotides in 5＇UTR,P1,P2,P3,3＇ UTR regions were 80. 3%-95. 7%,80. 8%-93. 9%,78. 6%-87. 7%,78. 1%-81. 2% and66. 7%-87. 5%,respectively; the similarities of nucleotides and amino acids of the VP1 region among these CV-B4 virus isolates from diabetic patients and hand,foot and mouth disease（HFMD） patients were 81. 2%-93. 2% and96. 1%-99. 3%,respectively. The largest difference was 28. 8% between Henan isolates and diabetes isolates,and6. 8% was the smallest difference with HFMD isolates. Conclusions The VP1 nucleotide homology of CV-B4 isolates from the same diseases is higher.