Development and evaluation of a rapid PCR method for the PowerPlex®S5 system for forensic DNA profiling

• A rapid PCR method for the PowerPlex®S5 system was developed and evaluated.
• Using a fast PCR approach, the amplification time was reduced by 70% (1 h).
• Sensitivity was similar to the standard protocol.
• Minor issues were observed with stutter percentages and peak height ratios.
• This rapid protocol is useful for screening large numbers of reference samples.

Forensic DNA profiling is a multi-step process taking approximately 10 h to complete. A reduction in the amount of time required for the amplification step would allow for faster human identification and increase laboratory throughput. The goal of this work was to optimize and evaluate a rapid PCR method for the PowerPlex®S5 system for forensic DNA profiling. By pairing fast chemistries with a fast thermal cycler, we were able to reduce the amplification time by 70% (1 h). Sensitivity and heterozygous peak height ratios were comparable between the fast and standard protocols. However, there was a notable decrease (5%) in peak height ratio at the D18S51 locus with the fast cycling method. An increase in average mean stutter for combined loci of 2.6% was observed in profiles amplified using the fast protocol compared to the standard system. Our results suggest that with further optimization and validation the fast protocol can be used to replace the standard amplification conditions.