中国临床保健杂志Issue(1)：63-67,5.DOI:10.3969/J.issn.1672-6790.2014.01.023
人乳腺癌特异性基因1 mRNA实时荧光定量逆转录-聚合酶链反应检测试剂盒的建立及应用
Establishment and application of a detection kit for human breast cancer-specific gene 1 mRNA by real-time ;quantitative RT-PCR
摘要
Abstract
Objective This study was to establish a quantitative method for human breast cancer-specific gene 1(Breast cancer-specific Gene 1,BCSG1)mRNA by real-time RT-PCR.Its application was further evaluated by detecting circulation tumor cells in the peripheral blood of breast cancer patients .Method The concentration of each component in the detection kit was optimized;the stability,sensitivity and specificity were determined .The expression level of BCSG1 mRNA was detected in the samples of peripheral blood .Results Optimized primers and probes were obtained,and setting standards were successfully prepared .Through establishing the positive and negative control ma-terials,the specificity,sensitivity,accuracy and quality control were determined for the detection kit .The specificity and accuracy were both more than 95%,and the sensitivity was of 100 copies/ml( whole peripheral blood ) .The detec-tion results of BCSG1 mRNA in peripheral blood samples showed that all the samples were positive for 12 breast canc-er patients clinically diagnosed with metastasis;and 12 samples were positive and other 9 samples were negative among 21 breast cancer patients without clinically diagnosed with metastasis or not ;all the samples were negative for 15 pa-tients with benign breast disease;and all the samples were negative for 10 healthy control .Conclusion The quantita-tive RT-PCR assay for BCSG1 mRNA could be used for the detection of circulating tumor cells in peripheral blood of breast cancer patients ,which would be valuable for the diagnosis and clinical-decision making for breast cancer patients .关键词
乳腺肿瘤/试剂盒,诊断/RNA,信使/逆转录聚合酶链反应Key words
Breast neoplasms/Reagent kits,diagnostic/RNA,messenger/Reverse transcriptase polymerase chain reaction分类
医药卫生引用本文复制引用
程民,沈国栋,卞庚,徐婷娟,徐维平,沈干,胡世莲..人乳腺癌特异性基因1 mRNA实时荧光定量逆转录-聚合酶链反应检测试剂盒的建立及应用[J].中国临床保健杂志,2014,(1):63-67,5.基金项目
国家自然科学基金面上项目（81071808）；安徽省国际科技合作项目（10080703037）；安徽省科技攻关项目（12010402126）；安徽省肿瘤免疫营养诊疗产业创新团队资助 （）
