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用反转录套式PCR(RT -PCR)法从陕西地区 1例NANB型肝炎患者血清中扩增出部分E区的cDNA片段 ,克隆于PGEM -T载体中用全自动序列分析仪进行双向测序 ,测得的序列以及已发表的同一分离株 5′NRC、C区序列 ,分别同Genbank中注册的 3 6株已知基因型的HCV相应区段的序列比较 ,构建种系发生树。结果 5′NCR区相应序列构建的种系发生树表明该分离株为 2型 ,而用C区和E区构建的种系发生树均表明该分离株为 2a型。提示该分离株为 2a型 ;对同一病原体来说 ,其基因区段的变异度不同 ,则种系发生树分析的结果不完全相同 ,变异度大的结果较准确。
A partial cDNA fragment of E region was amplified by reverse transcription polymerase chain reaction (RT-PCR) from one patient with NANB hepatitis in Shaanxi province. The cDNA fragment was cloned in PGEM-T vector and sequenced by full automatic sequence analyzer , The measured sequences and published sequences of the 5 ’NRC and C regions of the same isolate were compared with those of the corresponding 3 6 known genotypes registered in Genbank to construct the phylogenetic tree. Results The phylogenetic tree constructed with the corresponding sequences of 5’NCR showed that this isolate was type 2 whereas the phylogenetic tree constructed with both regions C and E showed that this isolate was type 2a. Suggesting that the isolated strain is type 2a. For the same pathogen, the degree of variation of the gene segments is different, and the results of the phylogenetic tree analysis are not exactly the same, and the result of the large variation degree is more accurate.