深圳市流行的麻疹野病毒的血清学和分子生物学研究

为探讨深圳市麻疹病毒的基因型别和特征 ,对深圳市专科医院 2 0 0 0～ 2 0 0 2年 2 2例临床诊断为麻疹病例的咽拭子、尿液标本提取核糖核酸 (RNA) ,通过逆转录 聚合酶链反应 (RT PCR)扩增麻疹病毒血凝素 (H)基因序列 ,并克隆到pMD18 T载体进行序列测定 ;分析这些麻疹病毒株和GeneBank中麻疹病毒各基因型代表株的序列同源性。采集病人血清 ,检测IgM抗体。结果显示 :2 2例麻疹病人的 2 2份血清中 ,有 16份IgM抗体阳性 ;2 0份咽拭子标本中 ,均检测到相应的条带 ;2 0份尿液标本中有 18份检测到相应的条带。采用RT PCR一次扩增的产物 ,检测的灵敏度即达到 0 0 0 1TCID50 。 19株麻疹病毒H基因之间的同源性为 96 2 %～ 10 0 0 % ,和H1基因型代表株相比同源性为 96 4 %～ 99 3%。根据分析结果 ,麻疹病人咽拭子标本的RT PCR阳性率高于尿液标本。该方法可用于麻疹病原学快速辅助诊断 ,特别是对出疹初期就诊的临床符合病例 ,但IgM抗体阴性的病例可进一步确诊。深圳市麻疹流行株属于H1基因型。

Study on Serology and Molecular Biology of Wild Measles Viruses Circulating in Shenzhen City

To explore the genetic c haracteristics and genotype of mea sles viruses circulating in Shenzh en city,extract RNA from throat sw abs and urine of 22 clinical diagn osed measles patients during 2000- 2002 by RT-PCR method to amplify t he gene sequence of measles virus hemagglutinin, and the PCR products were cloned into PMD 18-T carrier to test the sequence and analyze t he homogeneity of the sequence bet ween these measles virus and the r epresentative strains with each ge notype in GeneBank.Collect the serum of the pa tients and test IgM antibody.16 of 22 serum samples from 22 patients were IgM positive;all of 20 throat swabs were detected the correspond ing region.Using the product amplif ied by RT-PCR,the sensitivity of t he test achieved 0.001TCID 50 .H gen es homogeneity among the 19 strain s of measles virus was 96.2%-100%. Compared with the representative s train with H1 genotype,the homogen eity was 96.4%-99.3%.The results s howed that RT-PCR positive rate of the t hroat swabs was higher than that o f the urine for measles patients.T his method may help diagnose the p athogen of measles rapidly,especia lly to those patients with rash in the initial clinical stage but wit h negative measles IgM.The genotyp e of measles viruses circulation i n Shenzhen were belonged to clade H 1.