| 曾真,李沛涛,邬若楠,曹亚.一种简易基因水平鉴定HBV亚型的方法[J].Chinese journal of Epidemiology,1996,17(6):356-359 |
| 一种简易基因水平鉴定HBV亚型的方法 |
| A Simplified Method for Detection of Genomic Subtyps of HBV S-gene |
| Received:April 11, 1996 Revised:May 10, 1996 |
| DOI： |
| KeyWord: 基因 乙型肝炎病毒 |
| English Key Word: Gene HBV |
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| Abstract: |
| HBV S基因513bp的PCR扩增产物,用限制性内切酶Msp Ⅰ和BamH Ⅰ消化。adr亚型基因片段,用Msp Ⅰ可将其切割成为两个分别为320bp和193bp的片段;ayw亚型可被Msp Ⅰ消化切割成为326bp和187bp的两个片段;ayw的HBV扩增产物,也可用限制性内切酶BamH Ⅰ切割成295bp和218bp两条片段;adw亚型S基因无Msp Ⅰ的酶切位点,不被切割;adw、adr亚型HBV的S基因扩增产物无限制性内切酶BamH Ⅰ的酶切位点,不被切割。酶切后的产物用琼脂糖凝胶电泳,电脉谱型与设计所预期的一致。各HBV亚型的S基因RFLP谱型在电泳中各异,容易鉴别,且酶切后的DNA片段清晰可见便于观察。该法为HBV基因分型和分子流行病学调查提供了一种新的可靠手段。 |
| English Abstract: |
| When the 513bp amplified fragments of HBV S-gene, known subtypes, were digested with Msp Ⅰ and BamH Ⅰ, the following phenomena appeared:adr subtypic 513bp was fragmentized to 320bp and 193bp by Msp Ⅰ; ayw subtype was digested and fragmentized to 295bp and 218bp by BamH Ⅰ;ayw could also be fragmentized to become 326bp and 187bp digested by Msp Ⅰ. Howerev, adw subtype was not digested by Msp Ⅰ and BamH Ⅰ nor adr by BamH Ⅰ. The experimental patterns in agarose gel eleetrophoresis were tallied with the expected patterns. Each HBV subtypes gave unique size of digestive fragments which was easy to observe and distinguish on agarose gel. This study provided a new classification scheme for HBV-DNA and could be used for epidemiological investigation of HBV infection. |
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