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Proceedings Paper

Combining isothermal rolling circle amplification and electrochemiluminescence for highly sensitive point mutation detection
Author(s): Qiang Su; Xiaoming Zhou
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Paper Abstract

Many pathogenic and genetic diseases are associated with changes in the sequence of particular genes. We describe here a rapid and highly efficient assay for the detection of point mutation. This method is a combination of isothermal rolling circle amplification (RCA) and high sensitive electrochemluminescence (ECL) detection. In the design, a circular template generated by ligation upon the recognition of a point mutation on DNA targets was amplified isothermally by the Phi29 polymerase using a biotinylated primer. The elongation products were hybridized with tris (bipyridine) ruthenium (TBR)-tagged probes and detected in a magnetic bead based ECL platform, indicating the mutation occurrence. P53 was chosen as a model for the identification of this method. The method allowed sensitive determination of the P53 mutation from wild-type and mutant samples. The main advantage of RCA-ECL is that it can be performed under isothermal conditions and avoids the generation of false-positive results. Furthermore, ECL provides a faster, more sensitive, and economical option to currently available electrophoresis-based methods.

Paper Details

Date Published: 6 March 2009
PDF: 8 pages
Proc. SPIE 7280, Seventh International Conference on Photonics and Imaging in Biology and Medicine, 72801U (6 March 2009); doi: 10.1117/12.821576
Show Author Affiliations
Qiang Su, South China Normal Univ. (China)
Xiaoming Zhou, 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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