柯萨奇病毒B组4型河南分离株VP1区基因特征分析

VP1 characteristics of coxsakievirus B4 isolated in Henan province

目的 了解河南省手足口病病原柯萨奇病毒B组4型（CoxB4） VP1区基因特征,并比较不同疾病来源CoxB4病毒分离株VP1区异同.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;通过设计CoxB4病毒VP1区特异性引物,对8株病毒进行PCR扩增,用生物学软件ATGC进行序列拼接,ClustalX1.83进行序列比对,BioEdit剪齐,MEGA4.0进行同源性分析和构建系统进化树.结果 CoxB4河南分离株VP1区长852 bp,与GenBank公布的其他分离株核苷酸相似性为80.3％ ～97.0％,氨基酸相似性96.1％～100％;与糖尿病患者分离株E2核苷酸差异最大为19.7％,与手足口病分离株差异最小为3.0％.系统进化树显示CoxB4河南分离株属于Genotype Ⅴ型,手足口病分离株与脑膜炎分离株遗传距离较近,分布在一簇中,糖尿病患者分离株在另外一簇.结论 河南地区手足口病感染CoxB4病毒存在Genotype Ⅴ型传播和流行.

Objective To analyze the VP1 region characteristics of coxsakievirus B4 （CoxB4） strain isolated from patients with hand,foot and mouth disease （HFMD） in Henan province and to compare the similarity and difference among these viruses isolated from different diseases.Methods The 134 samples were tested by reverse-transcription polymerase chain reaction （RT-PCR） and cell-culture-based isolation of CoxB4.The VP1 of 8 CoxB4 isolated was amplified using specific primers.The sequences were spliced,edited,analyzed and mapped into phylogenetic tree by bioinformatics software ATGC,lustalx1.83,BioEdit and MEGA4.0,respectively.Results The VP1 region length of Henan CoxB4 isolates were 852 bp.Compared with other CoxB4 strains released in GeneBank,the similarities of nucleotide and amino acid is 80.3％-97.0％,96.1％-100％.The largest difference was 19.7％ between Henan isolates and diabetes isolates,and 3.0％ was the smallest difference with HFMD isolates.Phylogenetic tree analysis showed that Henan CoxB4 isolates belonged to Genotype Ⅴ,and the isolates from HFMD were close to those of meningitis,distributed in a cluster.These isolates from diabetic patients were in another cluster.Conclusions There wcre Genotype Ⅴ spreading and circulating of CoxB4 that infected HFMD patients in Henan.