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.

[1]  Douglas H Ubelaker,et al.  The use of SEM/EDS analysis to distinguish dental and osseus tissue from other materials. , 2002, Journal of forensic sciences.

[2]  Norah Rudin,et al.  Quality Assurance Standards for Forensic DNA Testing Laboratories , 2001 .

[3]  Bingcheng Lin,et al.  Determination of SARS‐coronavirus by a microfluidic chip system , 2004, Electrophoresis.

[4]  R S Foote,et al.  PCR amplification and analysis of simple sequence length polymorphisms in mouse DNA using a single microchip device. , 2000, Analytical biochemistry.

[5]  Peter M Vallone,et al.  Allele frequencies for 15 autosomal STR loci on U.S. Caucasian, African American, and Hispanic populations. , 2003, Journal of forensic sciences.

[6]  R. Piercy,et al.  Further validation of a multiplex STR system for use in routine forensic identity testing. , 1996, Forensic science international.

[7]  D N Stivers,et al.  The utility of short tandem repeat loci beyond human identification: Implications for development of new DNA typing systems , 1999, Electrophoresis.

[8]  Richard A Mathies,et al.  Rapid and High‐Throughput Forensic Short Tandem Repeat Typing Using a 96‐Lane Microfabricated Capillary Array Electrophoresis Microdevice * , 2006, Journal of forensic sciences.

[9]  Richard A Mathies,et al.  A Forensic Laboratory Tests the Berkeley Microfabricated Capillary Array Electrophoresis Device * , 2008, Journal of forensic sciences.

[10]  S. Lutz-Bonengel,et al.  Low-volume amplification on chemically structured chips using the PowerPlex16 DNA amplification kit , 2005, International Journal of Legal Medicine.

[11]  J. Mennell,et al.  The future of forensic and crime scene science. Part I. A UK forensic science user and provider perspective. , 2006, Forensic science international.

[12]  James P. Landers,et al.  Developments toward a complete micro-total analysis system for Duchenne muscular dystrophy diagnosis , 2003 .

[13]  Richard A Mathies,et al.  Microchip bioprocessor for integrated nanovolume sample purification and DNA sequencing. , 2002, Analytical chemistry.

[14]  William H. Grover,et al.  Monolithic membrane valves and diaphragm pumps for practical large-scale integration into glass microfluidic devices , 2003 .

[15]  Z. Hugh Fan,et al.  Integrating polymerase chain reaction, valving, and electrophoresis in a plastic device for bacterial detection. , 2003, Analytical chemistry.

[16]  Igor L. Medintz,et al.  High‐performance genetic analysis using microfabricated capillary array electrophoresis microplates , 2001, Electrophoresis.

[17]  Richard A Mathies,et al.  Inline injection microdevice for attomole-scale sanger DNA sequencing. , 2007, Analytical chemistry.

[18]  Jerome P Ferrance,et al.  Microchip‐Based Cell Lysis and DNA Extraction from Sperm Cells for Application to Forensic Analysis , 2006, Journal of forensic sciences.

[19]  Eric Buel,et al.  Forensic DNA typing by capillary electrophoresis using the ABI Prism 310 and 3100 genetic analyzers for STR analysis , 2004, Electrophoresis.

[20]  A. Manz,et al.  Miniaturized total chemical analysis systems: A novel concept for chemical sensing , 1990 .

[21]  R. Mathies,et al.  Fully integrated PCR-capillary electrophoresis microsystem for DNA analysis. , 2001, Lab on a chip.

[22]  Igor L. Medintz,et al.  Genotyping energy-transfer-cassette-labeled short-tandem-repeat amplicons with capillary array electrophoresis microchannel plates. , 2001, Clinical chemistry.

[23]  Brian N. Johnson,et al.  An integrated nanoliter DNA analysis device. , 1998, Science.

[24]  A. deMello Control and detection of chemical reactions in microfluidic systems , 2006, Nature.

[25]  R. Mathies,et al.  Multichannel reverse transcription-polymerase chain reaction microdevice for rapid gene expression and biomarker analysis. , 2006, Analytical chemistry.

[26]  Brian McKenna,et al.  A high‐performance multilane microdevice system designed for the DNA forensics laboratory , 2004, Electrophoresis.

[27]  J Buckleton,et al.  An investigation of the rigor of interpretation rules for STRs derived from less than 100 pg of DNA. , 2000, Forensic science international.

[28]  Thomas N. Chiesl,et al.  Ultrafast DNA sequencing on a microchip by a hybrid separation mechanism that gives 600 bases in 6.5 minutes , 2008, Proceedings of the National Academy of Sciences.

[29]  James P Landers,et al.  Forensic DNA Analysis on Microfluidic Devices: A Review , 2007, Journal of forensic sciences.

[30]  Richard A Mathies,et al.  Multichannel PCR-CE microdevice for genetic analysis. , 2006, Analytical chemistry.

[31]  Peng Liu,et al.  Integrated portable polymerase chain reaction-capillary electrophoresis microsystem for rapid forensic short tandem repeat typing. , 2007, Analytical chemistry.

[32]  Richard A Mathies,et al.  Microfabricated bioprocessor for integrated nanoliter-scale Sanger DNA sequencing. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[33]  Z Hugh Fan,et al.  Integrating polymerase chain reaction, valving, and electrophoresis in a plastic device for bacterial detection. , 2003, Analytical chemistry.

[34]  Susan A Greenspoon,et al.  Validation and implementation of the PowerPlex 16 BIO System STR multiplex for forensic casework. , 2004, Journal of forensic sciences.

[35]  P. Gill,et al.  Encoded evidence: DNA in forensic analysis , 2004, Nature Reviews Genetics.

[36]  J R Scherer,et al.  Integrated portable genetic analysis microsystem for pathogen/infectious disease detection. , 2004, Analytical chemistry.

[37]  S. Jacobson,et al.  Multiple sample PCR amplification and electrophoretic analysis on a microchip. , 1998, Analytical chemistry.

[38]  Benjamin E. Krenke,et al.  Validation of a 16-locus fluorescent multiplex system. , 2002, Journal of forensic sciences.

[39]  John W Bond,et al.  Value of DNA Evidence in Detecting Crime , 2007, Journal of forensic sciences.