Combining isothermal rolling circle amplification and electrochemiluminescence for highly sensitive point mutation detection

Qiang Su; Xiaoming Zhou

doi:10.1117/12.821576

6 March 2009 Combining isothermal rolling circle amplification and electrochemiluminescence for highly sensitive point mutation detection

Qiang Su, Xiaoming Zhou

Author Affiliations +

Proceedings Volume 7280, Seventh International Conference on Photonics and Imaging in Biology and Medicine; 72801U (2009) https://doi.org/10.1117/12.821576
Event: Photonics and Optoelectronics Meetings, 2008, Wuhan, China

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.

Citation Download Citation

Qiang Su and Xiaoming Zhou "Combining isothermal rolling circle amplification and electrochemiluminescence for highly sensitive point mutation detection", Proc. SPIE 7280, Seventh International Conference on Photonics and Imaging in Biology and Medicine, 72801U (6 March 2009); https://doi.org/10.1117/12.821576

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