Carcinogenesis, Teratogenesis & Mutagenesis ›› 1999, Vol. 11 ›› Issue (5): 213-216.doi: 10.3969/j.issn.1004-616x.1999.05.001

    Next Articles

QUANTITATIVE DETECTION OF TELOMERASE ACTIVITY BY COMBINATION OF TRAP METHOD AND SCINTILLATION COUNT ASSAY

Wu Chengqiu , Chen Wen , Zhang Qiao , et al   

  1. 1Department of Heal th Toxicology , School of Public Heal th , S un Yat2sen University of Medical Sciences , Guang Zhou 　510089 , 2 Department of Hygiene , Heng Yang Medical College , Heng Yang , Hu Nan 　 421001
  • Received:1900-01-01 Revised:1900-01-01 Online:1999-09-30 Published:1999-09-30

PDF (PC)

2732

Abstract: This study developed a semi2quantitative method of telomerase activity. The method was based on the combination of TRAP assay and scintillation count assay (3 H2dCTP incorporation) . It was used to quantify telomerase activity in CNE2 cells and some tissue specimens. RNase2pret reated or heat2t reated cells were used as cont rols. The result s demonst rated that telomerase activity measured by this method was positive in CNE2 cells , and it could be clearly detected with as few as 10 cells. There was a linear correlation between the radioactive count and the telomerase activity. The telomerase activity of some tissue specimens were accordant with reported data. Telomerase activity of RNase2pret reated or heatt reated cells was negative , their radioactive counts were almost the same as lysis buffer cont rol. The variations within group and between groups were 11. 68 % and 20.99 % , respectively. This method was f ree of PAGE and radioautography , and hence simple and fast . The results could be obtained with one day. It showed high sensitivity , good specificity and repeatability.

Key words: Telomerase, Quantitative Analysis, Telomeric Repeat Amplification Protocol, Scintillation Count Assay