Clinical Chemistry
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Clinical Chemistry 48: 1016-1020, 2002;
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(Clinical Chemistry. 2002;48:1016-1020.)
© 2002 American Association for Clinical Chemistry, Inc.

Bioluminescent Method for Detecting Telomerase Activity

Shun-Qing Xu1a, Min He1, Hong-Ping Yu1, Xiao-Yang Wang2, Xiang-Lin Tan1, Bin Lu1, Xi Sun1, Yi-Kai Zhou1, Qun-Feng Yao1, Yong-Jun Xu1 and Zhi-Ren Zhang1

1 National Laboratory of Biomedical Photonics, Institute of Environmental Medicine, Tongji Medical College of Huazhong University of Science and Technology, 13 Hangkong Rd., Wuhan 430030, The People’s Republic of China

2 Institute of Respiratory Disease, Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology, 1095 Jiefang Ave., Wuhan 430030, The People’s Republic of China

aAuthor for correspondence. Fax 86-27-83657705; e-mail shunqing{at}public wh.hb.cn.

Background: Telomerase is a promising biomarker in cancer diagnosis and therapy. The elongation of telomeric repeats catalyzed by telomerase is accompanied by release of six PPi for each TTAGGG repeat (1 pmol PPi/310 pg telomeric repeats). We developed a novel method to measure telomerase activity by use of an enzymatic luminometric PPi assay (ELIPA).

Methods: Extracts of cell lines and tissues were incubated with primer at 30 °C for 30 min. Released PPi was converted to ATP by sulfurylase, and ATP was detected by a luciferase bioluminescence system. The ELIPA results were compared with results obtained with the conventional telomeric repeat amplification (TRAP)-ELISA in 42 lung carcinoma tissues and 27 control tissues without malignancy.

Results: The lower detection limits of ELIPA and TRAP-ELISA were 5 and 10 cells, respectively. The within-run imprecision (CV) of ELIPA was <=12%. When compared with TRAP-ELISA, the correlation coefficient (r) was 0.79. When we used the cutoff value from ROC analysis to distinguish malignant and nonmalignant tissues, the sensitivity and specificity of ELIPA were 83% and 96%, respectively, whereas the sensitivity and specificity of TRAP-ELISA were 71% and 96%, respectively.

Conclusion: ELIPA is a simple and sensitive homogeneous method to quantify telomerase activity.




The following articles in journals at HighWire Press have cited this article:


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B. Lumbreras-Lacarra, J. M. Ramos-Rincon, and I. Hernandez-Aguado
Methodology in Diagnostic Laboratory Test Research in Clinical Chemistry and Clinical Chemistry and Laboratory Medicine
Clin. Chem., March 1, 2004; 50(3): 530 - 536.
[Abstract] [Full Text] [PDF]

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J. Grimm, J M. Perez, L. Josephson, and R. Weissleder
Novel Nanosensors for Rapid Analysis of Telomerase Activity
Cancer Res., January 15, 2004; 64(2): 639 - 643.
[Abstract] [Full Text] [PDF]


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