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Proceedings Paper

Sensitive detection of point mutation by electrochemiluminescence and DNA ligase-based assay
Author(s): Huijuan Zhou; Baoyan Wu
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Paper Abstract

The technology of single-base mutation detection plays an increasingly important role in diagnosis and prognosis of genetic-based diseases. Here we reported a new method for the analysis of point mutations in genomic DNA through the integration of allele-specific oligonucleotide ligation assay (OLA) with magnetic beads-based electrochemiluminescence (ECL) detection scheme. In this assay the tris(bipyridine) ruthenium (TBR) labeled probe and the biotinylated probe are designed to perfectly complementary to the mutant target, thus a ligation can be generated between those two probes by Taq DNA Ligase in the presence of mutant target. If there is an allele mismatch, the ligation does not take place. The ligation products are then captured onto streptavidin-coated paramagnetic beads, and detected by measuring the ECL signal of the TBR label. Results showed that the new method held a low detection limit down to 10 fmol and was successfully applied in the identification of point mutations from ASTC-α-1, PANC-1 and normal cell lines in codon 273 of TP53 oncogene. In summary, this method provides a sensitive, cost-effective and easy operation approach for point mutation detection.

Paper Details

Date Published: 6 March 2009
PDF: 9 pages
Proc. SPIE 7280, Seventh International Conference on Photonics and Imaging in Biology and Medicine, 72802O (6 March 2009); doi: 10.1117/12.821581
Show Author Affiliations
Huijuan Zhou, South China Normal Univ. (China)
Baoyan Wu, South China Normal Univ. (China)


Published in SPIE Proceedings Vol. 7280:
Seventh International Conference on Photonics and Imaging in Biology and Medicine
Qingming Luo; Lihong V. Wang; Valery V. Tuchin, Editor(s)

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