| 摘要: |
| ［摘要］目的 建立一种快速、准确、特异的定量检测ACACB基因rs2268388多态性的方法．方法 针对ACACB基因序列设计引物和Taqman探针，探索实时荧光定量PCR技术检测ACACB基因rs2268388多态性的最佳条件，设立不同浓度，并检测其灵敏度和特异性．结果 PCR反应预变性时间为10 min，退火、延伸时间为1 min时扩增产物效果理想． Taqman实时荧光定量PCR探针及引物浓度 0.5 μL （ 0.2 μM），TaqMan Genotyping Master Mix 5 μL ，dH2O 2.5 μL总体系为10 μL时扩增产物效果理想并节约了实验费用，检测特异性良好．结论 优化Taqman探针法的实验条件能够灵敏、特异、稳定地对ACACB基因rs2268388多态性进行定量检测，并有效节约了实验成本. |
| 关键词: ［关键词］Taqman实时荧光定量PCR ACACB基因 特异性 敏感性 |
| DOI： |
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| 基金项目:［基金项目］国家自然科学基金资助项目（30760087，81160104）；云南省自然科学基金资助项目（2004C0066M） |
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| Experimental Conditions of Detecting rs2268388 Polymorphism of ACACB Gene |
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TANG Jun-ting1,2
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1.（1）Dept. of diabetes;2）Dept. of Laboratory，The 1st Affiliated Hospital of Kunming Medical University， Kunming Yunnan 650032，China）
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| Abstract: |
| ［Abstract］Objective To establish a rapid and specific quantitative PCR assay for detecting of ACACB gene rs2268388 polymorphism. Methods The primers and Taqman probe targeted at signature sequence of ACACB gene rs2268388 polymorphism were designed and applied to real time PCR assays. The different concentrations were set up， and the sensitivity and specificity were evaluated. Results Initial denaturation time of PCR was 10 min， annealing and extending time were 1 min， probe concentration of Taqman PCR was 0.5 μL（0.2 μM），TaqMan Genotyping Master Mix was 5 μL， dH2O was 2.5 μL，and total volume was 10 μL. At this condition，the result of ACACB gene rs2268388 polymorphism showed the high specificity of the reaction. Conclusion The assay was sensitive and specific for rapid identification and quantitative detection of ACACB gene rs2268388 polymorphism. It will be an effective method for the surveillance and diagnosis of ACACB gene rs2268388 polymorphism. |
| Key words: ［Key words］Taqman polymerance chain reaction ACACB gene Sensitive Specificity |