摘要
目的 建立多重逆转录 聚合酶链反应 (RT PCR)方法半定量检测肝癌多药耐药(MDR)基因mRNA表达。方法 在对二重RT PCR扩增 5种MDR基因mRNA动力学观察及建立RT PCR标准曲线的基础上 ,建立六重RT PCR方法。结果 六重RT PCR与单重或二重RT PCR扩增体系在特异性和扩增效率等方面相同 ,实验重复性较好。结论 该方法对于检测肝癌组织和肝癌细胞MDR基因mRNA表达具有特异、灵敏、简便、快速 (6h)、所需标本量小 (5mg)及半定量等优点 ,能快速同时分析大量样本 ,具有临床应用前景。
Objective To establish multiple RT PCR semi quantitative detection system to test MDR gene mRNA expression.Methods Using duplicate RT PCR as method through a process of amplification to become a multi component RT PCR.Based on observation of 5 MDR gene's mRNA kinetic effect studies and the setting of RT PCR standard curves, a six complex RT PCR method was established.Results Six complex RT PCR system was identical to simple and duplicated RT PCR in sensitivity and specificity, and easily to be repeated.Conclusion This given method was sensitive, specific, and had the advantages of speed in performance (within 6 h),only small quantity of specimen (50 mg) and semi quantitation when used to detect the MDR gene mRNA expression in primary liver cancer tissue and cell line. It could also be applied to process a batch of samples simultaneously in one setting.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2002年第6期598-600,共3页
Chinese Journal of Experimental Surgery
