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为研究ECHO13病毒中国分离株的分子特征及其与世界其它分离株之间的基因关系,对1998年、2000年从中国福建省分离到2株ECHO13病毒进行基因序列对比分析。2株病毒分别命名为Fujian98-1和Fujian00-1,用逆转录-聚合酶链反应(RT-PCR)扩增出VP1蛋白编码基因全长861个核苷酸片段并进行序列测定,将2株E CHO13病毒的VP1序列与所有已发表的ECHO13病毒VP1基因全长进行同源性比较。结果显示,福建分离株之间核苷酸同源性为79 6%,氨基酸同源性为93 4%;与遗传距离最近的法国CF1089-91(AJ537604)毒株的核苷酸同源性分别为80%和88%,与代表株DelCarmen的核苷酸同源性分别为75 8%和77 9%。通过VP1基因分析,福建2株病毒均属于ECHO13病毒,与血清中和试验鉴定结果一致。下载所有已发表的ECHO13病毒VP1序列并构建进化树,发现福建2株病毒分属不同的分枝,提示这2株病毒来自不同的病毒传播链。进一步分析发现,整个E CHO13病毒可划分为3个不同的基因型:A、B和C基因型。福建Fujian98-1和Fujian00-1分别被划分在基因型B和C中,各基因型之间的核苷酸差异均大于20%。为验证该分型方法,将26株来自不同国家和时间的ECHO13病毒和2株福建分离ECHO13病毒部分VP1基因序列进行对比分析,建立进化树。结果显示,所有ECHO13病毒被分在A、B和C3个基因
In order to study the molecular characteristics of ECHO13 isolate in China and its genetic relationship with other isolates in the world, two ECHO13 isolates were isolated from Fujian Province in China in 1998 and 2000 respectively. The two viruses were named Fujian98-1 and Fujian00-1 respectively. The full-length 861 nucleotide fragment of the VP1 protein gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) The VP1 sequence of E CHO13 virus was compared for homology with the full length of all published VP1 gene of ECHO13 virus. The results showed that the nucleotide homology between Fujian isolates was 79 6% and the amino acid homology was 93 4%. The nucleotide homology with the French CF1089-91 (AJ537604) strain with the closest genetic distance was 80% and 88%, respectively. Nucleotide homology with the representative strain DelCarmen was 75.8% and 77.9%, respectively. By VP1 gene analysis, Fujian 2 viruses belong to ECHO13 virus, and serum identification test results are consistent. Download all published VP1 sequences of ECHO13 virus and construct phylogenetic tree. It was found that the two viruses in Fujian belong to different branches, suggesting that the two viruses originated from different virus transmission chains. Further analysis found that the entire E CHO13 virus can be divided into three different genotypes: A, B and C genotypes. Fujian Fujian98-1 and Fujian00-1 were divided into genotypes B and C, respectively, and the nucleotide differences among all genotypes were greater than 20%. In order to validate the typing method, we compared the VP1 gene sequences of 26 ECHO13 isolates from different countries and regions and 2 Fujian isolates of ECHO13 virus to establish phylogenetic tree. The results showed that all ECHO13 viruses were subdivided into A, B and C3 genes