HCV 定量 RT-PCR 竞争性参比标准的构建  

CONSTRUCTION OF A PLASMID WHICH CAN TRANSCRIBE A MUTANT HCV RNA AS THE COMPETITIVE TEMPLATE IN QUANTITATIVE RT PCR

作　　者：吕凌[1] 王斌[1] 

机构地区：[1]中山医科大学分子医学研究中心

出　　处：《中山医科大学学报》1997年第2期151-153,共3页Academic Journal of Sun Yat-sen University of Medical Sciences

基　　金：广东省自然科学基金

摘　　要：将克隆的ＨＣＶ５’ＮＣＲ序列反向插入ｐＳＰ７２的ＥｃｏＲＶ切点，构建一种能转录ＨＣＶ－ＲＮＡ的质粒ｐＳｎｃ－２。以ＲＴ－ＰＣＲ从抗ＨＥＶＩｇＭ阳性病人血浆中扩增一段２３８ｂｐ的ＨＣＶ－ｃＤＮＡ，反向插入ｐＳｎｃ－２中克隆的５’ＮＣＲ序列的ＮｃｏⅠ切点，构建插入突变型ＨＣＶ－ｃＤＮＡ重组体ｐＳｎｃ－ｅ。应用限制性酶切和ＰＣＲ对这两种重组质粒进行了鉴定，为ＨＣＶ－ＲＮＡ的定量ＰＣＲ提供有用的内参照模板。A 238 bp HEV cDNA was amplified from plasma of a patient positive to anti HEV IgM. It was inserted into the NcoⅠ site of HCV 5'NCR sequence cloned into the vector pSP72. A plasmid pSnc e was recombinated that can transcribe a mutated HCV RNA serving as the competitive RNA template for quantitative RT PCR assay. The pSnc e was determined by PCR and restrictive site analysis.

关 键 词：克隆 分子 聚合酶链反应 遗传学 丙型肝炎病毒 

分 类 号：R512.630.3[医药卫生—内科学] R394[医药卫生—临床医学]