聚合酶链反应检测肿瘤组织的端粒酶活性

目的：比较聚合酶链反应－微孔板杂交法（PCR－MPH）和聚合酶链反应－聚丙烯酰胺凝胶电泳法（PCR－PAGE）检测肿瘤组织端粒酶活性的临床价值。方法：PCR－MPH以地高辛标记端粒重复片段特异的探讨与PCR变性产物杂交，经酶底物显色，通过测定吸光度判断结果；PCR－PAGE方法直接用聚丙烯酰胺凝胶电泳法分离PCR扩增端粒酶的合成产物，经银染色，分析端粒酶活性。结果：PCR－MPH方法能准确、特异地检测出肿瘤组织的端粒酶活性，其灵敏度比PCR－PAGE法高100倍，结论：两种方法均可用于临床检测，只是PCR－MPH方法更简单、便一些。

Polymerase chain reaction in determining telomerase activity in tumor tissues

Objectives:To detect telomerase activity in various tumor tissues by PCR PAGE and PCR MPH. Methods:We used telomerase PCR MPH,an assay in which a digoxigenin labeled telomerase repeat specific probe was hybridized to the PCR product. After revelation with an anti digoxin AKP conjugates, the amount of hybridized probe was determined by optical reading. Telomerase activity was analyzed by PAGE. Results:Telomerase activity was determined by PCR MPH accurately and specifically in various tumor tissues, its sensitivity was one hundred times higher than that of PCR PAGE. Conclusion:PCR MPH is ideal for analyzing telomerase activity in various tumor tissues. It is simple, easy and suitable for clinical application.