title: Rapid microfluidic analysis of a Y-STR multiplex for screening of forensic samples.
abstract: In this paper, we demonstrate a rapid analysis procedure for use with a small set of rapidly mutating Y chromosomal short tandem repeat (Y-STR) loci that combines both rapid polymerase chain reaction (PCR) and microfluidic separation elements. The procedure involves a high-speed polymerase and a rapid cycling protocol to permit PCR amplification in 16 min. The resultant amplified sample is next analysed using a short 1.8-cm microfluidic electrophoresis system that permits a four-locus Y-STR genotype to be produced in 80 s. The entire procedure takes less than 25 min from sample collection to result. This paper describes the rapid amplification protocol as well as studies of the reproducibility and sensitivity of the procedure and its optimisation. The amplification process utilises a small high-speed thermocycler, microfluidic device and compact laptop, making it portable and potentially useful for rapid, inexpensive on-site genotyping. The four loci used for the multiplex were selected due to their rapid mutation rates and should proved useful in preliminary screening of samples and suspects. Overall, this technique provides a method for rapid sample screening of suspect and crime scene samples in forensic casework. Graphical abstract ᅟ.
authors.affiliation: Department of Chemistry and Biochemistry and International Forensic Research Institute, Florida International University, University Park, Miami, FL, 33199, USA.
authors.forename: Georgiana
authors.initials: G
authors.lastname: Gibson-Daw
authors.affiliation: School of Chemical Sciences, University of Auckland, 23 Symonds Street, Building 302, Auckland, 1010, New Zealand.
authors.forename: Patricia
authors.initials: P
authors.lastname: Albani
authors.affiliation: Agilent Technologies, Hewlett-Packard-Straße 8, 76337, Waldbronn, Germany.
authors.forename: Marcus
authors.initials: M
authors.lastname: Gassmann
authors.affiliation: Department of Chemistry and Biochemistry and International Forensic Research Institute, Florida International University, University Park, Miami, FL, 33199, USA. mccordb@fiu.edu.
authors.forename: Bruce
authors.initials: B
authors.lastname: McCord
mesh_terms: D041322:Chromosomes, Human, Y; D004247:DNA / Q000302:isolation & purification; D019075:Electrophoresis, Capillary; D005260:Female; D044707:Forensic Sciences; D006801:Humans; D057230:Limit of Detection; D008297:Male; D044085:Microfluidics; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results
chemical_list: D004247:DNA
keywords: Forensic; Genotyping; Microfluidic; Rapid PCR; Y short tandem repeat