摘要
目的建立一种快速检测CYP2D6*10基因多态性的方法。方法基于荧光定量PCR平台,采用Taqman探针分型技术结合扩增阻滞突变聚合酶链式反应(ARMS-PCR)技术,建立一种适应多种类型临床样本(抗凝全血、石蜡组织切片等)的快速检测CYP2D6*10基因的方法。使用多种类型的临床样本及不同浓度的基因组DNA,对检测体系的准确性、灵敏度进行验证。结果该研究建立的检测体系适用抗凝全血、石蜡包埋组织等多种类型的临床样本,基因组DNA检测范围为30~30000 copy/μL,检测结果不受同源基因干扰,且与Sanger直接测序法检测结果一致率为100%。结论该研究建立的CYP2D6*10荧光定量PCR检测体系准确性好、灵敏度高,具有广泛的应用价值。
Objective To establish a rapid detection method for CYP2D6*10 gene polymorphism.Methods Based on the fluorescent quantitative PCR platform,the Taqman probe typing method combined with amplification refractory mutation system technology was used to establish a rapid assay for detection of CYP2D6*10 suitable for various types of clinical samples(anticoagulated whole blood,paraffin tissue sections,etc).Various types of clinical samples and different concentrations of genomic DNA were used to verify the accuracy and sensitivity of the detection system.Results The detection system established in this study was suitable for anticoagulated whole blood,paraffin-embedded tissue and other types of clinical samples.The range of genomic DNA detection was 30-30000 copy/μL,and the detection results were not interfered by homologous genes,and the consistency rate with the Sanger sequencing result was 100%.Conclusion The established CYP2D6*10 fluorescent quantitative PCR detection system exhibits high accuracy and sensitivity,and has a wide range of application values.
作者
吴建元
章柏钰
蔡君龙
汪再兴
刘凤麟
曾超
黄建英
WU Jianyuan;ZHANG Baiyu;CAI Junlong;WANG Zaixing;LIU Fenglin;ZENG Chao;HUANG Jianying(Clinical Trial Center,Zhongnan Hospital of Wuhan University,Wuhan,Hubei 430071,China;Wuhan YZY Medical Science and Technology Co.,Ltd.,Wuhan,Hubei 430075,China)
出处
《国际检验医学杂志》
CAS
2022年第14期1713-1720,共8页
International Journal of Laboratory Medicine
基金
湖北省卫生健康委员会科研项目面上项目(WJ2021M169)
武汉大学中南医院2021年度医学科技创新平台支撑项目(lcyf202107)。
