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通过聚合酶链反应(PCR)检测法快速诊断耐药基因。

Rapid diagnosis of drug-resistant genes by PCR assay.

作者信息

Funato T

机构信息

Department of Clinical and Laboratory Medicine, Tohoku University School of Medicine, Sendai.

出版信息

Rinsho Byori. 1995 Jun;43(6):535-9.

PMID:7602796
Abstract

This report concerns the utility of the reverse transcription-polymerase chain reaction (RT-PCR) and quantitative PCR (QPCR) assay to detect the drug-resistance of related genes. The expression of some drug-resistance genes was compared with the sensitivity and resistance-acquired cancer cell lines to anti-cancer drugs by Northern blot analysis and PCR assay. The resistance cell lines exhibited an enhanced expression of multi-drug resistance (MDR-1), thymidylate synthase (TS), c-fos and DNA polymerase beta genes. Then these genes that expressed mRNA were quantitated using RT-PCR. The expression of the genes was dependent on their sensitivity (IC50) to anti-cancer drugs. Additionally, the QPCR assay has been developed as a rapid method for the expression of drug-resistance genes and applied to the PCR products amplified by the RT-PCR. Thus the QPCR assay for the expression of genes will allow rapid detection of the drug-resistance to chemotherapy in human cancers.

摘要

本报告涉及逆转录-聚合酶链反应(RT-PCR)和定量PCR(QPCR)检测相关基因耐药性的实用性。通过Northern印迹分析和PCR检测,将一些耐药基因的表达与对抗癌药物敏感和获得耐药性的癌细胞系进行了比较。耐药细胞系表现出多药耐药(MDR-1)、胸苷酸合成酶(TS)、c-fos和DNA聚合酶β基因的表达增强。然后使用RT-PCR对这些表达mRNA的基因进行定量。这些基因的表达取决于它们对抗癌药物的敏感性(IC50)。此外,已开发出QPCR检测作为一种快速检测耐药基因表达的方法,并应用于RT-PCR扩增的PCR产物。因此,用于基因表达的QPCR检测将能够快速检测人类癌症中对化疗的耐药性。

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