Interrogation of short tandem repeats using fluorescent probes and melting curve analysis: a step towards rapid DNA identity screening.
French DJ., Howard RL., Gale N., Brown T., McDowell DG., Debenham PG.
Current forensic DNA profiling methods rely on the analysis of samples at specialised laboratories with an average turnaround time of several days. The ability to rapidly determine a partial profile of short tandem repeats at the point-of-arrest would be of great benefit to police forces around the world, for example enabling a suspect to be rapidly included or excluded from an investigation. We have developed a homogeneous PCR method for the interrogation of STR loci utilising fluorescent oligonucleotide probes and melting curve analysis. Alleles of the D18S51, TH01 and D8S1179 loci were differentiated and identified on the basis of target length and probe melting temperature. Assay performance was evaluated by comparing melting peak data with the AmpFlSTR SGM Plus system. The method is compatible with direct analysis of unpurified buccal swab samples, enabling a partial STR profile to be generated within 1h.