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目的分离并鉴定北京市输入性D9基因型麻疹毒株。方法使用Vero/SLAM细胞,对可疑麻疹输入性病例的咽拭子和尿液标本进行麻疹毒株的分离培养。用反转录-聚合酶链反应扩增麻疹病毒核蛋白(N)基因羧基末端676个核苷酸片段,对扩增产物进行核苷酸序列测定和分析,并以羧基末端450个核苷酸片段构建基因亲缘关系树,进行遗传距离及核苷酸同源性分析。结果该病毒分离株BJCY13026-2和世界卫生组织D9基因型代表株Victoria.AUS(维多利亚.澳大利亚)12.99在基因亲缘性关系树上同属一个分支,核苷酸同源性为95.8%,氨基酸同源性为96%;和其他23个基因型代表株的核苷酸和氨基酸同源性分别在88.1%~95.6%和90.7%~96.7%。和中国大陆目前所使用的麻疹疫苗株沪191相比对,其核苷酸和氨基酸同源性分别为91.1%和90.0%;和中国目前流行的麻疹病毒绝对优势本土基因型H1a基因型代表株相比对,其核苷酸和氨基酸同源性分别为89.4%和91.3%。结论该输入性病例的病毒分离株为麻疹病毒D9基因型。
Objective To isolate and identify Beijing imported D9 genotype measles strains. Methods Vero / SLAM cells were used to isolate and isolate measles isolates from throat swabs and urine samples of suspected measles cases. The 676 nucleotides of the carboxyl terminus of the measles virus nucleoprotein (N) gene were amplified by reverse transcription - polymerase chain reaction (PCR). The amplified product was sequenced and analyzed. The nucleotide sequence of 450 nucleotides Fragment construction of genetic relationship tree, genetic distance and nucleotide homology analysis. Results The virus isolate BJCY13026-2 and WHO D9 genotype representative strain Victoria. AUS (Victoria, Australia) 12.99 belong to the same branch of the gene relationship tree, with 95.8% nucleotide homology, amino acid homology The nucleotide and amino acid homologies of the representative strains of other 23 genotypes were 88.1% -95.6% and 90.7% -96.7%, respectively. Compared with the measles vaccine strain Shanghai 191 currently used in mainland China, the nucleotide and amino acid homologies were 91.1% and 90.0%, respectively; and the Chinese predominant H1N1 genotype representative strain of measles virus In comparison, their nucleotide and amino acid homologies were 89.4% and 91.3%, respectively. Conclusion The virus isolate of this imported case is the D9 genotype of measles virus.