Improved forensic DNA analysis through the use of alternative DNA polymerases and statistical modeling of DNA profiles.
DNA evidence, linking perpetrators to crime scenes, is central to many legal proceedings. However, DNA samples from crime scenes often contain PCR-inhibitory substances, which may generate blank or incomplete DNA profiles. Extensive DNA purification can be required to rid the sample of these inhibitors, although these procedures increase the risk of DNA loss. Most forensic laboratories use commercial DNA amplification kits (e.g., AmpFlSTR SGM Plus) with the DNA polymerase AmpliTaq Gold as the gold standard. Here, we show that alternative DNA polymerase-buffer systems can improve the quality of forensic DNA analysis and efficiently circumvent PCR inhibition in crime scene samples, without additional sample preparation. DNA profiles from 20 of 32 totally or partially inhibited crime scene saliva samples were significantly improved using Bio-X-Act Short, ExTaq Hot Start, or PicoMaxx High Fidelity instead of AmpliTaq Gold. A statistical model for unbiased quality control of forensic DNA profiles was developed to quantify the results. Our study demonstrates the importance of adjusting the chemistry of the PCR to enhance forensic DNA analysis and diagnostic PCR, providing an alternative to laborious sample preparation protocols.
Hedman
Johannes
J
Department of Applied Microbiology, Lund University, Lund, Sweden.
Nordgaard
Anders
A
Rasmusson
Birgitta
B
Ansell
Ricky
R
Rådström
Peter
P
D004247:DNA / Q000032:analysis / Q000737:chemistry* / Q000235:genetics*; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry*; D003627:Data Interpretation, Statistical; D053803:Forensic Genetics / Q000379:methods*; D006801:Humans; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods*
D004247:DNA; D004259:DNA-Directed DNA Polymerase