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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
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, 0(7): 748-751
作者姓名:苏全喜  李婉仪  张成  熊符  洪铭范  刘爱群  田素娟
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;
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)~~
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。

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
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, 0(7): 748-751
Authors:SU Quan-xi  LI Wan-yi  ZHANG Cheng  XIONG Fu  HONG Ming-fan  LIU Ai-qun  TIAN Su-juan
Affiliation: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...
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....
Keywords:Muscular Dystrophy  Facioscapulohumeral  Real-Time Quantitative Pcr  Rnase H Enzyme Activity  
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