东莞市2008年麻疹病毒基因型分析

An analysis on genotype of wild type measles virus circulated in Dongguan City in 2008

目的：为了掌握东莞市2008年流行的麻疹病毒的基因型别和核蛋白基因变异情况。方法：采集东莞市2008年麻疹疫情中病人的血清、咽拭子、尿液等标本。对血清IgM抗体检测阳性的标本进行病毒分离,采用VERO-SLAM细胞进行培养,对分离到的病毒株提取核糖核酸（RNA）,通过逆转录-聚合酶链反应（RT-PCR）扩增麻疹病毒N基因序列,对扩增产物进行测序,取病毒株碳末端450bp核苷酸序列进行分析。结果：共分离到11株麻疹病毒株,均属于H1基因型H1 a亚型。11株毒株间的核苷酸同源性为98%~100%,氨基酸同源性为96.7%~100%,11株毒株间,2、4、5、7、8号的序列相同,9、10、11号的序列相同。各毒株与Ch ina93-2毒株同源性最高,为98.7%~99.3%,与Ch ina94-7毒株同源性为96.7%~97.3%,与其它H1亚型的同源性则为97.1%~98.2%。结论：东莞市2008年麻疹病毒流行株均属于H1基因型H1 a亚型,与中国其它地区的流行情况一致。暂无新型及传入性的其它亚型出现。

Objective：To know the genotype of wild type measles viruses circulating in 2008 in Dongguan City and the mutation situation of the nucleoprotein gene.Methods：To collect the serum,throat swabs or urine specimens from measles patients in outbreaks and isolate the viruses by inoculating the throat swabs or urine specimens to Vero-Slam cell line.To extract the RNA from the viruses.The COOH-terminal of nucleoprotein gene was amplified by RT-PCR,the expected length of the fragment was 594 bp.PCR products were purified,sequenced and the 450 bp COOH-terminal of nucleoprotein gene fragment was analyzed.Results：11 viruses were isolated from patients in outbreaks,Phylogenetic analysis of the N gene sequences showed that all of the isolates belonged to H1 genotype H1a subtype.The homology of nucleotides sequence and predicted amino acids sequence of these viruses ranged from 98% to 100% and 96.7% to 100% respectively,the number 2,4,5,7,8 have the same sequence,the number 9,10,11 have the same sequence.all the viruses have 98.7%~99.3%,96.7%~97.3% and 97.1%~98.2% homology of nucleotides sequence with China 93_2 virus,China94_7 virus and other H1 genotype virus respectively.Conclusion：11 measles virus circulateing in Dongguan City were H1 genotype H1a subtype,it is in accordance with the situation of china.No any other genotype virus or new genotype virus emerged in dongguan city.