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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