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This work concerns the identification of the allelic sequences of the DQB1gene of the human leukocyte antigen system conferring susceptibility to thedevelopment of insulin-dependent diabetes mellitus (IDDM) in DNA samples withno need of PCR amplification. Our method is based on the time-resolved analysisof a F\"orster energy-transfer mechanism that occurs in a dual-labeledfluorescent probe specific for the base sequence of the allelic variant ofinterest. Such an oligonucleotide probe is labeled, at the two ends, by a pairof chromophores that operate as donor and acceptor in a F\"orster resonantenergy-transfer. The donor fluorescence is quenched with an efficiency that isstrongly dependent on the donor-to-acceptor distance, hence on theconfiguration of the probe after hybridization with the DNA containing or notthe selected allelic sequence. By time-correlated single-photon counting,performed with an excitation/detection system endowed with 30-ps resolution, wemeasure the time-resolved fluorescence decay of the donor and discriminate, bymeans of the decay time value, the DNA bearing the allele conferringsusceptibility to IDDM from the DNAs bearing any other sequence in the sameregion of the DQB1 gene.

Polymorphic gene conferring susceptibility to insulin-dependent diabetes mellitus typed by ps-resolved FRET on nonamplified genomic DNA