TRAP-silver staining, a highly sensitive assay for measuring telomerase activity in tumor tissue and cell lines.
نویسندگان
چکیده
Measurement of telomerase activity in clinically obtained tumor samples may provide important information for use as both a diagnostic marker and a prognostic indicator for patient outcome. In order to evaluate telomerase activity in tumor tissue without radiolabeling the product, we developed a simple telomeric repeat amplification protocol-silver-staining assay that is less time-consuming, is safe and requires minimal equipment. In addition, we determined the sensitivity of the silver-staining method by using extracts of telomerase-positive thyroid carcinoma cell lines which were serially diluted from 5,000 to 10 cells. Telomerase activity was also assayed in 19 thyroid tumors, 2 normal controls and 27 bone marrow aspirates. The results indicate that the technique permits the detection of telomerase activity from 5000 to as few as 10 cells. We propose that it could be immediately applicable in many laboratories due to the minimal amount of equipment required.
منابع مشابه
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BACKGROUND Telomerase is a ribonucleoprotein enzyme associated with immortalization and transformation of human cells. The telomeric repeat amplification protocol (TRAP) is widely used for the detection of telomerase activity. The TRAP method, although highly sensitive and specific because it includes PCR amplification, is laborious and does not provide precise quantitative information. METHO...
متن کاملA non-isotopic method for the detection of telomerase activity in tumour tissues: TRAP-silver staining assay.
Telomerase activity has been observed in a high proportion of specimens from a wide variety of human malignant tumours, but not in non-neoplastic tissues. Measurement of telomerase activity is a possible diagnostic maker for malignant tumours. A non-isotopic telomeric repeat amplification protocol (TRAP) silver staining method for the detection of telomerase activity in human bone tumour tissue...
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عنوان ژورنال:
- Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas
دوره 35 1 شماره
صفحات -
تاریخ انتشار 2002