Real-time forensic DNA analysis at a crime scene using a portable microchip analyzer.
An integrated lab-on-a-chip system has been developed and successfully utilized for real-time forensic short tandem repeat (STR) analysis. The microdevice comprises a 160-nL polymerase chain reaction reactor with an on-chip heater and a temperature sensor for thermal cycling, microvalves for fluidic manipulation, a co-injector for sizing standard injection, and a 7-cm-long separation channel for capillary electrophoretic analysis. A 9-plex autosomal STR typing system consisting of amelogenin and eight combined DNA index system (CODIS) core STR loci has been constructed and optimized for this real-time human identification study. Reproducible STR profiles of control DNA samples are obtained in 2h and 30min with <or=0.8bp allele typing accuracy. The minimal amount of DNA required for a complete DNA profile is 100 copies. To critically evaluate the capabilities of our portable microsystem as well as its compatibility with crime scene investigation processes, real-time STR analyses were carried out at a mock crime scene prepared by the Palm Beach County Sheriff's Office (PBSO). Blood stain sample collection, DNA extraction, and STR analyses on the portable microsystem were conducted in the field, and a successful "mock" CODIS hit was generated on the suspect's sample within 6h. This demonstration of on-site STR analysis establishes the feasibility of real-time DNA typing to identify the contributor of probative biological evidence at a crime scene and for real-time human identification.
Liu
Peng
P
UCSF/UC Berkeley Joint Graduate Group in Bioengineering, University of California, Berkeley, CA 94720, USA.
Yeung
Stephanie H I
SH
Crenshaw
Karin A
KA
Crouse
Cecelia A
CA
Scherer
James R
JR
Mathies
Richard A
RA
D003415:Crime; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D016172:DNA Fingerprinting / Q000379:methods*; D004867:Equipment Design; D005240:Feasibility Studies; D053803:Forensic Genetics / Q000379:methods*; D005784:Gene Amplification; D006801:Humans; D018895:Microsatellite Repeats / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction
D004247:DNA