Evaluation of an in vitro telomeric repeat amplification protocol assay to detect telomerase activity in canine urine

Objective-To evaluate the usefulness of a PCR-based telomeric repeat amplification protocol (TRAP) assay for detecting telomerase activity in cells from a canine transitional cell carcinoma (TCC) cell line and ultimately in the urine of dogs with TCC Animals-11 dogs with histologic or cytologic evidence of TCC 10 dogs with benign lower urinary tract disease and 9 healthy dogs Procedures-Telomerase activity was initially evaluated in cells from canine TCC (K9TCC) and telomerase-negative (WI-38) cell lines Following assay optimization telomerase stability was evaluated at various storage durations and temperatures Urine samples were then obtained prospectively from study dogs Results-Telomerase activity was detected in the K9TCC cell line The TRAP assay detected telomerase activity in as few as 10 K9TCC cells alone and as low as 2% of a total cell population in K9TCC and WI-38 mixing experiments A loss of telomerase activity was detected with increasing urine storage durations at various temperatures Telomerase activity was clearly detected in samples collected from 10 of 11 dogs with TCC 2 of 10 dogs with benign lower urinary tract disease and none of the 9 healthy dogs Conclusions and Clinical Relevance-The TRAP based assay detected telomerase activity in the canine TCC cell line and revealed that the telomerase ribonucleoprotein complex was inherently unstable at various storage durations and conditions Telomerase activity was also detectable in urine samples obtained from dogs with TCC which suggested the TRAP assay may be useful in diagnosing TCC in dogs (Am J Vet Res 2010 71 1468-1474)