| 面肩肱型肌营养不良实时荧光定量PCR诊断方法的影响因素分析 |
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| 引用本文： | 苏全喜,李婉仪,张成,熊符,洪铭范,刘爱群,田素娟.面肩肱型肌营养不良实时荧光定量PCR诊断方法的影响因素分析[J].军医进修学院学报,2011(7):748-751. |
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| 作者姓名： | 苏全喜 李婉仪 张成 熊符 洪铭范 刘爱群 田素娟 |
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| 作者单位： | 广东药学院附属第一医院神经内科;东莞市人民医院神经内科;中山大学附属第一医院神经科;南方医科大学基础医学院医学遗传学教研室;广东药学院生命科学与生物制药学院; |
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| 基金项目： | 国家自然科学基金项目(30870851); 广东省自然科学基金面上项目(8151008018000002)~~ |
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| 摘 要： | 目的分析探讨面肩肱型肌营养不良（FSHD）实时荧光定量聚合酶链反应（FQ-PCR）诊断方法的影响因素。方法通过改变镁离子终浓度、PCR扩增中退火温度、退火时间或延伸时间、RNaseH酶用量和活性等，确定FQ-PCR诊断FSHD的最佳反应条件。结果退火温度55℃、时间20s、镁离子终浓度5．OmM、RNaseH酶用量IOOU、RNaseH酶在95℃下灭活8min为最佳反应条件，其中RNaseH的活性选择对FSHD诊断的特异性影响较大。结论通过降低RNaseH酶的活性可提高诊断的特异性，最终确立了FSHD的FQ-PCR基因诊断最佳反应条件。
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| 关 键 词： | 肌营养不良 面肩肱型 实时荧光定量PCR RNase H酶活性 |
Factors influencing diagnosis of facioscapulohumeral muscular dystrophy by real-time fluorescent quantitative PCR |
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| SU Quan-xi,LI Wan-yi,ZHANG Cheng,XIONG Fu,HONG Ming-fan,LIU Ai-qun,TIAN Su-juan.Factors influencing diagnosis of facioscapulohumeral muscular dystrophy by real-time fluorescent quantitative PCR[J].Academic Journal of Pla Postgraduate Medical School,2011(7):748-751. |
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| Authors: | SU Quan-xi LI Wan-yi ZHANG Cheng XIONG Fu HONG Ming-fan LIU Ai-qun TIAN Su-juan |
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| Institution: | SU Quan-xi1,LI Wan-yi2,ZHANG Cheng3,XIONG Fu4,HONG Ming-fan1,LIU Ai-qun1,TIAN Su-juan5 1Department of Neurology,First Affiliated Hospital of Guangdong Pharmaceutical University,Guangzhou 510080,Guangdong Province,China,2Department of Neurology,Dongguan People's Hospital,Dongguan 523018,3Department of Neurology,First Affiliated Hospital of Sun Yat-Sen University,4Department of Medical Genetics,School of Basic Medical Sciences,Southern Medical... |
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| Abstract: | Objective To analyze the factors influencing diagnosis of facioscapulohumeral muscular dystrophy(FSHD) by real-time fluorescent quantitative polymerase chain reaction(FQ-PCR).Methods Optimal reaction conditions of FQ-PCR for the diagnosis of FSHD were established by changing the annealing temperature,annealing and elongation time of FQ-PCR,as well as the Mg2+ concentration,the amount of RNase H used and its activity.Results The optimal reaction conditions of FQ-PCR for the diagnosis of FSHD were as follows.... |
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| Keywords: | Muscular Dystrophy Facioscapulohumeral Real-Time Quantitative Pcr Rnase H Enzyme Activity |
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