Detection of known mutations for medical diagnostics by FRET spectroscopy.

A rapid, simple and low-cost method for the detection of known mutations in DNA oligonucleotide in a biothreat agent, Bacillus anthracis, has been reported. The technique is based on fluorescence resonance energy transfer (FRET), that utilizes a cationic conjugated polymer and a PNA probe labeled with Fluorescein dye (PNAC*). When the PNA probe is hybridized with a complementary target ssDNA and its mutated sequences separately, the energy transfer from polymer to PNAC*/ssDNA complex decreases with increasing number of mutations. It means that the efficiency of FRET or the degree of hybridization depends on the extent of mutations in the DNA sequence. The method is sensitive enough to detect upto 4 bases mismatch. We have, thus, explored a possible application of fluorescence-based technology for medical diagnostics.