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基于聚合酶链反应的酶免疫测定法定量端粒酶活性:在结直肠癌中的应用

Polymerase chain reaction-based enzyme immunoassay for quantitation of telomerase activity: application to colorectal cancers.

作者信息

Cheng A J, Tang R, Wang J Y, Chang J T, Wang T C

机构信息

Department of Medical Technology, Chang Gung University, Tao-Yuan, Taiwan.

出版信息

Jpn J Cancer Res. 1999 Mar;90(3):280-5. doi: 10.1111/j.1349-7006.1999.tb00745.x.

Abstract

Telomerase is a specialized reverse transcriptase that synthesizes telomeric sequences onto human chromosomal ends. It appears to be present in the majority of primary human cancer tissues, and may have potential as a universal tumor marker. In this report, we describe a sensitive, non-radioactive, polymerase chain reaction (PCR)-based enzyme immunoassay (EIA) for the quantitation of telomerase activity in human cells. This PCR-EIA is convenient and can be easily completed within 3 h. The correlation coefficient between the results of PCR-EIA and the conventional telomeric repeat amplification protocol (TRAP) method, as measured on 4 different cell lines, was over 0.98. Evaluation of this method for clinical application was conducted with tissues obtained from patients with colorectal cancers and the results were compared with those of the conventional TRAP method. Our data indicate that telomerase activities measured by conventional TRAP and PCR-EIA are highly correlated, and we suggest that the PCR-EIA method can substitute for conventional TRAP.

摘要

端粒酶是一种特殊的逆转录酶,可将端粒序列合成到人类染色体末端。它似乎存在于大多数原发性人类癌症组织中,可能具有作为通用肿瘤标志物的潜力。在本报告中,我们描述了一种基于聚合酶链反应(PCR)的灵敏、非放射性酶免疫测定法(EIA),用于定量人类细胞中的端粒酶活性。这种PCR-EIA方法简便,可在3小时内轻松完成。在4种不同细胞系上测量,PCR-EIA结果与传统端粒重复序列扩增法(TRAP)之间的相关系数超过0.98。用从结直肠癌患者获取的组织对该方法进行临床应用评估,并将结果与传统TRAP方法的结果进行比较。我们的数据表明,传统TRAP法和PCR-EIA法测得的端粒酶活性高度相关,我们建议PCR-EIA法可替代传统TRAP法。

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