Quantitative detection of telomerase activity by real-time TRAP assay in the body fluids of cancer patients.
Real-time TRAP assay was developed to improve the sensitivity and quantitative detection of telomerase activity in the body fluids of cancer patients. The sensitivity and clinical significance of the real-time TRAP assay was investigated. Real-time PCR protocol was modified by using ACX primer and SYBR green mixture from the process of TS primer extension. Real-time TRAP showed high correlation (r2=0.843, p=0.001) and sensitivity (25 times higher) compared to conventional TRAP. Of 164 samples, there were 8 positives in cytology (4.9%), 7 (4.3%) using the conventional TRAP, and 41 (25%) using real-time TRAP. In cytology positive samples, real-time TRAP showed a higher positivity than conventional TRAP (75% vs 63%) suggesting a higher sensitivity in the body fluids. There was a tendency towards a longer progression-free duration in telomerase negative patients than in positive patients, as determined by conventional and real-time TRAP. The diagnostic interval between the first positivity documentation and clinical progression was short in the order of real-time TRAP, conventional TRAP and cytology. In conclusion, real-time TRAP assay can detect telomerase activity at an earlier phase of cancer progression and can replace conventional TRAP assay for detecting the telomerase activity in body fluids.
Shim
Woo Young
WY
Cancer Metastasis Research Center, Yonsei University College of Medicine, Shinchon-dong, Seoul, Korea.
Park
Kyu Hyun
KH
Jeung
Hei-Chul
HC
Kim
Yong Tae
YT
Kim
Tae Soo
TS
Hyung
Woo Jin
WJ
An
Sung Hwan
SH
Yang
Sang Hwa
SH
Noh
Sung Hoon
SH
Chung
Hyun Cheol
HC
Rha
Sun Young
SY
D001826:Body Fluids / Q000737:chemistry*; D006801:Humans; D009369:Neoplasms / Q000175:diagnosis* / Q000201:enzymology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D019098:Telomerase / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D014407:Tumor Cells, Cultured
D019098:Telomerase