Mr. Qiang Su Profile

Qiang Su

at South China Normal Univ

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Proceedings Article | 6 March 2009 Paper

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

Qiang Su, Xiaoming Zhou

Proceedings Volume 7280, 72801U (2009) https://doi.org/10.1117/12.821576

KEYWORDS: Magnetism, Target recognition, Electrodes, Target detection, Polymers, Biological research, Genetics, Life sciences, Pathogens, Ruthenium

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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.

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