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目的检测昆明地区麻疹病毒的优势基因型,为麻疹监测和防治提供依据。方法采集2008年昆明地区临床诊断为麻疹的30例患儿,其中男性18例,女性12例;年龄51天～3岁,0～1岁(≤1岁)的患儿19例,1～2岁(≤2岁)的患儿9例,2～3岁(≤3岁)的患儿2例,平均患病年龄15个月。采集急性传染期静脉血2 mL,应用酶联免疫吸附分析(ELISA)检测麻疹病毒血清IgM抗体;同时收集患儿的尿液标本,运用逆转录-聚合酶链反应(RT-PCR)扩增标本中所分离到的麻疹野病毒株核蛋白(N)基因羧基末端的543个核苷酸,对扩增产物进行核苷酸序列测定和分析,并与麻疹病毒基因库中的参考株进行比较。结果从30份麻疹患儿尿液标本中分离到7份麻疹野病毒阳性;出疹后6 d的尿液标本中仍可检出麻疹野病毒;将其4株麻疹野病毒的核蛋白(N)羧基末端的543个核苷酸与Genebank中的麻疹病毒的23个参考株进行基因亲缘性关系比较,与世界卫生组织(WHO)的H1、H2基因型参考株China 93-7、China 94-1所对应的核苷酸序列的遗传距离分别为0.031～0.047和0.332～0.398;而与H1a的遗传距离最近,为0.010～0.033;4株麻疹野病毒N基因羧基末端543个核苷酸的同源性为96.0%～98.9%;与H1标准株China 93-7和H1a标准株China 93-2的核苷酸同源性分别为95.9%～99.7%和96.9%～99.8%。结论2008年昆明地区所检测到的麻疹野病毒的优势基因型为H1a基因型。
Objective To detect the dominant genotypes of measles virus in Kunming area and provide evidence for measles surveillance and prevention. Methods Thirty children with measles were collected from Kunming in 2008, including 18 males and 12 females, 19 children aged 51 days to 3 years and children aged 0 to 1 years (≤1 years) There were 9 cases of children (≤2 years old) and 2 cases of children aged ≤3 years (≤3 years old) with an average age of 15 months. Collect 2 mL of venous blood in acute phase of infection and detect the serum IgM antibody of measles virus by enzyme-linked immunosorbent assay (ELISA). Urine samples from children were collected and amplified by RT-PCR The nucleotide sequence of the amplification product was determined and analyzed and compared with that of the reference strain in the measles virus gene pool. Results Seven samples of wild measles virus were isolated from urine samples of 30 measles children. Wild-type measles virus could still be detected in the urine specimens of 6 days after the measles eruption. The nucleoprotein (N ) Were compared with the 23 reference strains of measles virus in Genebank. The results showed that there was no significant relationship between the carboxy-terminal 543 nucleotides and the reference strains of H1 and H2 genotypes of the World Health Organization (WHO) China 93-7, China 94- 1 was 0.031 ~ 0.047 and 0.332 ~ 0.398, respectively, while the closest genetic distance to H1a was 0.010 ~ 0.033. The nucleotide sequence of 543 nucleotides of N gene of 4 wild measles virus The nucleotide homology was 96.9% -99.7% and 96.9% -99.8%, respectively, to H1 standard strain China 93-7 and H1a standard strain China 93-2. Conclusion The predominant genotype of wild measles virus detected in Kunming in 2008 was H1a genotype.