TRAP-ELISA法检测妇科恶性肿瘤组织中端粒酶活性

Quantitative detection of telomerase activity by TRAP-ELISA

目的 :探讨应用端粒重复序列扩增 (TRAP) 酶联免疫吸附测定法 (ELISA)定量检测妇科恶性肿瘤组织中的端粒酶活性的应用价值。方法 :采用TRAP ELISA法 ,对 36例宫颈癌、16例宫颈上皮内瘤样病变 (CIN)、2 5例非癌宫颈组织、2 5例卵巢上皮性肿瘤和 6例正常卵巢的新鲜组织及 41例宫颈脱落细胞进行端粒酶活性定量检测。结果 :宫颈组织端粒酶活性总阳性率 ,CIN 5 6 2 5 % (9/ 16 ) ,宫颈癌 91 6 7% (33/ 36 ) ,非癌宫颈组织 4% (1/ 2 5 ) ;宫颈脱落细胞 ,CIN 6 1 5 4% (8/ 13) ,宫颈癌 82 35 % (14/ 17) ,正常 0 % (0 / 11) ;卵巢组织端粒酶活性阳性率 ,良性卵巢上皮性肿瘤 2 5 % (2 / 8) ,交界性 6 6 6 7% (2 / 3) ,恶性 85 7% (12 / 14) ,正常卵巢 16 6 7% (1/ 6 )。恶性肿瘤与正常或良性病变中端粒酶活性的差异有显著性。结论 :妇科恶性肿瘤组织中端粒酶活性明显高于良性病变和正常组织。TRAP ELISA法较传统的TRAP法更为简便快速 ,有材料多样 ,可定量 ,特异性强、敏感性高 。

Aim:To investigate detective methodology of telomerase activity and its clinical application Methods: The telomerase activity was measured by telomere repeat amplification protocol (TRAP) and Enzyme Linked Immuno Sorbent Assay (ELISA), which amplicons containing biotiny TS primer were combined with microtiter plate coated streptavidin, then hybridized with DIG-labeled detection probe complementary to telomeric repeat sequences and finally combined with anti-digoxigenin-peroxidase antibody Results:Telomerase activity in tissues was detected in 9/16(5625%)CIN, 33/36 (9167%)invasive cervical cancer and 1/4 (25%)controls, in scrapings from 8/13(6154%) CIN, 14/17(8235%) invasive cervical cancer and 0/11(0%)normal controls, in 2 /8 benign(25%), 2 /3 borderline-malignant(667%), 12/ 14 malignant ovarian tumors(857%), and 1/6 cortex of normal ovaries(1667%) There was statistically significant difference between malignant tumor and benignIn or normal tissues Conclusion: The assay has no isotope contamination It provides a way to perform a higher sentive, simpler, more rapid and photometric enzyme immunoassay for the quantitative detection of telomerase activity than conventional protocol