Three-component Förster resonance energy transfer (FRET) has been used to obtain efficient FRET between the cationic conjugated polymer (CCP) as donor and 5-carboxy tetramethylrhodamine (TAMRA) dye as acceptor, by using an intermediate donor, fluorescein. In spite of the fact that there is enough overlap between the emission spectra of CCP and absorption spectra of TAMRA, the efficiency of FRET between CCP and TAMRA is poor. The reason for this is that while the Förster critical distance is not very sensitive to the overlap, the FRET efficiency is extremely sensitive to it. However, it is observed that the FRET efficiency between CCP and TAMRA improves considerably when fluorescein is introduced in the solution. The triple FRET so obtained can be used for deoxyribonucleic acid sequence detection in medical diagnostics because the fluorescence emission from TAMRA is pH-insensitive.

A reliable, fast, and low-cost biosensor for medical diagnostics using DNA sequence detection has been developed and tested for the detection of the bacterium "Bacillus anthracis." In this sensor, Poly [9,9-di (6,6 -N, N trimethylammonium) hexylfluorenyl-2, 7-diyl)-alt-co-(1,4-phenylene)] dibromide salt (PFP) has been taken as cationic conjugated polymer (CCP) and PNA attached with fluorescein dye (PNAC * ) as a probe. The basic principle of this sensor is that when a PNAC * probe is hybridized with a single strand DNA (ssDNA) having complementary sequence, Forster resonance energy transfer (FRET) may take place from PFP to the PNAC * /DNA complex. If the FRET is efficient, the photoluminescence from the PFP will be highly quenched and that from PNAC * will be enhanced. On the other hand, if the DNA sequence is noncomplementary to PNA, FRET will not occur.

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.

An improved strategy has been developed for detection of DNA sequence by using water-soluble cationic conjugated polymer (PFP)/single-strand (ss) DNA and peptide nucleic acid labeled with fluorescent dye (PNAC*), where an anionic surfactant (sodium dodecyl sulphate, SDS) system has been used to improve the sensitivity of the sensor. The method of detection is simple to use, fast and cost-effective. This method uses the phenomenon of Forester Resonance Energy Transfer (FRET). The detection sensitivity of the biosensor has been improved by about ten times by using the anionic surfactant. It is observed that the effect of surfactant is to increase the photoluminescence (PL) intensity of the PNAC* when the sequence of the DNA is complementary (to that of PNA probe). On the other hand when the two sequences are non-complementary, the PL intensity of the PNAC* is further reduced as compared to the case when surfactant was absent.