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Improvement in detecting telomerase activity using silica-based resin treatment: An experience of urine in bladder
N Yahata1, K Nagao, J H Ohyashiki
1First Department of Internal Medicine, Tokyo Medical University, Shinjuku-ku, Tokyo 160-0023, Japan.
Abstract:
To improve the telomeric repeat amplification protocol (TRAP) assay to detect telomerase activity using a small amount of sample, we used a resin-column to purify and to concentrate the TS extension DNA sequence. We used 14 samples of naturally voided urine (10 ml) from patients with bladder carcinoma and 9 urine samples from patients with non-malignant urological neoplasias. We used ethylenediamine tetraacetic acid (EDTA) to stabilize telomerase activity and resin treatment to concentrate TS-extended DNA and to exclude PCR inhibitor(s), and then performed extract-based fluorescence TRAP to detect telomerase activity. None of the urinary samples without resin-column treatment had detectable telomerase activity, whereas, in resin-column treated samples, 4/9 (44%) urine samples without EDTA and 9/14 (64%) with EDTA treatment had detectable telomerase activity. A combination of EDTA treatment and resin-column thus may be available to detect telomerase activity using a relatively small amount of secretion fluids, including exfoliated urinary cells.
