摘要目的 分析贵州2012—2015年风疹病毒分离株基因特征.方法 于2012—2015年贵州省麻疹网络实验室共采集390例疑似麻疹病例,并从中诊断出25例风疹病例,使用Vero/SLAM细胞进行病毒分离.经Real-time RT-PCR方法 鉴定为阳性的风疹病毒分离株,采用RT-PCR方法 扩增风疹病毒E1基因两个核苷酸片段(480 bp和633 bp),在对扩增产物进行序列测定和拼接后,基于基因定型靶基因(739 bp)进行基因特征分析.结果 25例疑似风疹病例中,19例为风疹暴发病例,6例为风疹散发病例;男性11例(44.0%),女性14例(56.0%),年龄为(12.3±3.9)岁;共分离到10株风疹病毒株, 7株为1E基因型,3株为2B基因型.7株1E基因型间核苷酸和氨基酸同源性分别为99.0%~100%和100%;3株2B基因型间核苷酸和氨基酸同源性分别为99.4%~100%和99.5%~100%.10株风疹病毒在E1糖蛋白基因Asn 177、Asn 209 N-型糖基化位点以及位于213~285 aa之间E1抗原表位未发生变异.其中,7株1E基因型在第338位氨基酸由亮氨酸变异为苯丙氨酸;2株2B基因型在第377位氨基酸由缬氨酸变异为丙氨酸.结论 2012—2015年在贵州传播流行的风疹病毒是由1E和2B基因型引起;10株风疹病毒株在核苷酸和氨基酸序列水平上高度保守,重要功能位点均未发生变异.

abstractsObjective To analyze the genetic characteristics of rubella virus isolated from 2012 to 2015 in Guizhou province. Methods A total of 390 cases of suspected measles were collected from Guizhou measles network laboratory from 2012 to 2015 and 25 cases of rubella cases were diagnosed. Rubella virus isolation was performed using Vero/SLAM cells. The presence of rubella viral RNA was detected using Real-time RT-PCR after RNA extraction from infected tissue culture cells. Fragments of 480 bp and 633 bp nucleotides of E1 genes of the isolates were amplified by RT-PCR and the PCR products were sequenced and spliced. The phylogenetic tree was conducted based on the 739 bp nucleotide sequences of E1 genes and gene characteristic analysis was performed. Results There were 19 cases of rubella outbreaks and 6 cases of rubella sporadic cases in 25 cases of suspected rubella cases.There were 11 males(44.0%)and 14 females(56.0%).The mean age and standard deviation were(12.3±3.9)years.A total of 10 rubella strains were isolated.The results of phylogenetic analysis showed that 7 strains of rubella virus isolates belonged to genotype 1E and the other belonged to genotype 2B. The nucleotide acid and amino acid homology among 7 strains 1E genotype were 99.0%-100% and 100% respectively.2B genotype of 3 strains of nucleotide and amino acid homology were 99.4%-100% and 99.5%-100% respectively.Ten strains of rubella virus were not mutated in the E1 glycoprotein gene, Asn 177 and Asn 209 N-type glycosylation sites and E1 antigen epitopes between 213 and 285aa.Among them, 7 strains of 1E genotype had a mutation from leucine to phenylalanine in 338 amino acid,2 strains of 2B genotype at 377 amino acids from valine to alanine. Conclusion Rubella virus epidemic was caused by 1E and 2B genotypes in Guizhou from 2012 to 2015.Ten strains of rubella virus were highly conserved in nucleotide and amino acid sequences and there was no variation of important functional sites.