NCJ Number
225051
Journal
Forensic Science International: Genetics Volume: 2 Issue: 4 Dated: September 2008 Pages: 333-339
Date Published
September 2008
Length
7 pages
Annotation
This paper reports on the development of a homogeneous PCR method for the interrogation of short tandem repeat (STR) DNA loci by using fluorescent oligonucleotide probes and melting curve analysis, which enables the production of a partial STR profile within 1 hour.
Abstract
This study demonstrates the proof-of-principle that melting curve analysis, using fluorescent oligonucleotide probes, will enable rapid interrogation of a small panel of STR loci outside of specialized laboratories. HyBeacon tests are compatible with unpurified saliva and buccal swab samples and may provide forensic DNA evidence within 1 hour by using a real-time PCR instrument. The combination of fluorescent probe and nonfluorescent blocker oligonucleotides enabled efficient detection and identification of D18S51, TH01, and D8S1179 repeat alleles. The D18S51 and D8S1179 assays clearly discriminated between the common alleles, comprising 11-21 and 8-17 repeats, respectively, with all Tm data within the defined acceptance criteria of the means + three standard deviations. All STR alleles may be reliably identified by melting peak Tm, with the exception of a 17/18+ D18S51 repeat combination, which can be determined visually. The generation of a partial STR profile at the point-of-screening is not intended to replace full forensic profiling methods, but will permit forensic DNA evidence to be rapidly produced before a sample could arrive at a specialized laboratory. The ability to provide a rapidly determined partial profile of short tandem repeats at the point-of-arrest would enable police to make a rapid determination of whether a suspect should be included or excluded from an investigation. Further work is required in order to reduce test complexity and ensure that assays are sufficiently accurate and robust for forensic analysis. 4 tables, 3 figures, and 19 references