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目的 了解甲型肝炎病毒 (HAV)在中国几个城市的基因型分布 ,为HAV的分子流行病学追踪调查提供方法和依据。方法 17株HAV代表株分别来自不同城市的甲型肝炎病人粪便或血清 ,病毒RNA经蛋白酶K消化、酚 /氯仿提取和乙醇沉淀后 ,以逆转录 套式聚合酶链反应 (RT PCR)扩增合成HAVVP1/ 2A交接区基因区 ,并进行直接核苷酸序列分析和差异比较。结果 VP1/ 2A交接区核苷酸序列分析表明 ,所有病毒株均从属于基因Ⅰ型 ;约 53%为ⅠB亚型 ,亚型间差异小于6 % ;约 4 7%为ⅠA亚型 ,亚型间差异小于 5 3% ;ⅠA与ⅠB亚型间的同源性为 88 7% - 92 3%。结论 中国流行或散发的HAV株可能有基因ⅠA、ⅠB亚型同时存在 ,流行病学相关的病毒株核苷酸序列相同或相近。
Objective To understand the genotype distribution of Hepatitis A virus (HAV) in several cities in China and provide a method and basis for molecular epidemiological investigation of HAV. Methods Seventeen HAV strains were from the excrement or serum of hepatitis A patients from different cities. The viral RNA was amplified by reverse transcription polymerase chain reaction (RT PCR) after protease K digestion, phenol / chloroform extraction and ethanol precipitation. The HAVVP1 / 2A junction gene region was synthesized, and direct nucleotide sequence analysis and difference comparison. Results Nucleotide sequence analysis of VP1 / 2A junction showed that all the strains belonged to genotype Ⅰ, about 53% were subtype ⅠB, the difference was less than 6% among subtypes, and 47% were subtype ⅠA and subtype The difference between ⅠA and ⅠB subtypes was 88 7% -92 3%. Conclusions There may exist both genotypes IA and IB in HAV strains that are prevalent or sporadic in China. The nucleotide sequences of epidemiologically related strains are the same or similar.