江西省赣州市2013—2016年麻疹病毒分离株N基因和H基因特征分析

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目的:分析赣州市流行的麻疹病毒的基因特征。方法:采集麻疹疑似病例咽拭子标本,应用荧光RT-PCR方法检测麻疹病毒核酸;用Vero/SLAM细胞对麻疹病毒核酸检测阳性标本进行病毒分离培养,并将麻疹病毒分离株进行核蛋白N基因和血凝素蛋白H基因扩增、测序;运用DNAStar、Bioedit和MEGA 5.0等分子生物学软件对N基因和H基因进行序列特征分析。结果:978份疑似麻疹病例咽拭子标本中,检出47份麻疹病毒核酸阳性,阳性率为4.81%,分离到25株毒株;扩增的13株毒株均为H1基因型的H1a亚型,与H1a基因型代表株(CHN/93/2)N基因核苷酸、氨基酸的同源性分别98.0%~98.5%、97.9%~99.6%,与H基因核苷酸、氨基酸的同源性分别为98.2%~98.9%、97.4%~98.5%。各分离株N基因与中国疫苗株Shanghai-191之间核苷酸、氨基酸的同源性分别为93.7%~95.2%、95.2%~96.8%;H基因与Shanghai-191之间核苷酸、氨基酸的同源性为94.2%~96.5%、94.2%~96.6%。结论:2013—2016年赣州市麻疹病毒流行株为H1a亚型,疫苗株对现流行株有保护性,但仍需持续监测麻疹病毒基因的变异情况。“,”Objective:To understand the genetic characterization of measles virus circulating in Ganzhou city.Methods:The throat swab specimens from suspected measles cases were collected and measles virus nucleic acid were detected by real-time RT-PCR. The Vero/SLAM cell line was used to isolate the virus strains from the positive samples. The nuclear protein N gene and hemagglutinin protein H gene of the virus were amplified and sequenced. The DNAStar, Bioedit and MEGA5 software were used to analyze the characters of N gene and H gene.Results:From the 978 specimens of suspected measles cases, 47 were tested positive of nucleic acids of measles virus with the positive rate of 4.81%. Twenty-five strains of measles virus were isolated. Thirteen strains were amplified and all were H1a subtype of the H1 genotype. The similarities of nucleotide and amino acid sequences of N gene with the representative H1a strains (CHN/93/2) were 98.0%-98.5% and 97.9%-99.6%, respectively while similarities of nucleotide and amino acid sequences of H gene were 98.2%-98.9% and 97.4%-98.5% , respectively. The similarities of N gene with the vaccine strain in China (Shanghai-191) were 93.7%-95.2% in nucleotides sequence and 95.2%-96.8% in amino acid sequence. For H gene, the similarities were 94.2%-96.5% in nucleotides sequence and 94.2%-96.6% in amino acid sequence.Conclusions:The circulating strains of measles virus in Ganzhou city from 2013 to 2016 was of the H1a subtype. The vaccine strain was protective against the current virus strain, but it was still necessary to continuously monitor the mutation of the measles virus gene.
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