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Alternative direct-to-amplification cell lysis techniques for forensically relevant non-sperm cells.

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A new method using Mammalian Protein Extraction Reagent (M-PER™) efficiently lyses non-sperm cells in sexual assault samples. This direct lysis technique speeds up DNA analysis by eliminating purification steps, aiding in reducing evidence backlogs.

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Area of Science:

  • Forensic Science
  • Molecular Biology
  • Biochemistry

Background:

  • Sexual assault evidence collection kits face significant backlogs due to lengthy laboratory DNA analysis.
  • Current DNA workflows require time-consuming differential lysis and complex mixture analysis.
  • A direct lysis method for sperm cells exists, but not for non-sperm cells.

Purpose of the Study:

  • To identify a rapid, cost-effective lysis method for non-sperm cells in sexual assault samples.
  • To find an alternative to traditional differential lysis that bypasses DNA purification steps.
  • To optimize DNA extraction from vaginal swabs for improved forensic analysis.

Main Methods:

  • Vaginal swab samples were lysed using various reagents, including alkaline buffers (25-200 mM NaOH), RIPA buffer, M-PER™, digitonin buffer, and urea/thiourea buffer.
  • Lysates were quantified using Quantifiler® Trio to assess lysis efficiency.
  • Short tandem repeat (STR) profiles were analyzed to evaluate the quality of DNA amplification.

Main Results:

  • The alkaline (25 mM NaOH) and M-PER™ methods effectively lysed vaginal epithelial cells with minimal sperm cell lysis.
  • M-PER™ demonstrated promising results in STR profile analysis, including high allele detection and good peak balance.
  • The M-PER™ method produced amplification-ready lysates without requiring a DNA purification step.

Conclusions:

  • M-PER™ is a viable alternative to traditional differential lysis for non-sperm cells in sexual assault investigations.
  • This method offers a faster and simpler approach to DNA analysis, potentially reducing evidence processing times.
  • Implementing M-PER™ could help alleviate the backlog of sexual assault evidence collection kits.