DD3基因实时荧光定量RT—PCR检测前列腺癌特异性方法的建立  

Establishing Real-time Fluorescent Quantitative Reverse Transcription Polymerase Chain Reaction Methods Specific for DD3 Genes to Detect Prostatic Carcinoma

作　　者：李娅[1] 岳乔红[1] 苏明权[1] 杨柳[1] 马越云[1] 郝晓柯[1] 

机构地区：[1]第四军医大学西京医院全军临床检验医学中心,西安710032

出　　处：《现代检验医学杂志》2009年第2期14-16,共3页Journal of Modern Laboratory Medicine

基　　金：第四军医大学创新基金（编号：20070608）.

摘　　要：目的建立DD3基因实时荧光定量PCR检测前列腺癌特异性方法，为前列腺癌的快速诊断提供分子诊断依据.方法根据Genebank中的DD3mRNA全序列，设计DD3基因的特异引物和探针，采用基因重组技术构建用于DD3基因检测的定量标准品;建立DD3基因的实时荧光定量方法.结果成功构建了用于DD3基因荧光定量PCR检测的标准重组质粒和DD3基因实时荧光定量PCR方法;经稳定性、特异度、重复性和敏感度实验方法学评价，DD3基因实时荧光定量PCR方法的最低检测限为1．64copy/ml，线性范围为1．64-1．64×10^12copy/ml.结论DD3基因实时荧光定量PCR检测前列腺癌特异性方法的建立，将为前列腺癌特异性诊断及其临床应用奠定方法学基础.Objective To establish real-time fluorescent quantitative RT-PCR methods specific for DD3 genes to detect prostatic carcinoma and supply molecular evidence for quick prostatic carcinoma diagnosis. Methods Specific primer and marked probes for DD3 gene were designed according to full DD3 mRNA sequence in Genbank. Quantitative standards for DD3 gene were set up with molecular recombination methods. To establish real-time fluorescent quantitative RT-PCR methods for DD3 genes. Results Standard reconstructed plasmid and DD3 genes-specific fluorescence quantitative RTPCR method were established successfully. After laboratory evaluation on stability,specificity,repeatability and sensitivity. The lowest limitation for detecting DD3 gene by real-time fluorescence quantitative PCR was 1.64 copy/ml. It＇s line scope ranged from 1.64 to 1.64×10^12 copy/ml. Conclusion The establishment of real-time fluorescent quantitative RT- PCR methods specific for DD3 genes to detect prostatic carcinoma will set up base for prostatic carcinoma diagnosis and potential clinical application.

关 键 词：DD3基因 实时荧光定量PCR 前列腺癌 

分 类 号：R730.45[医药卫生—肿瘤] Q503[医药卫生—临床医学]