Automating direct-to-PCR for disaster victim identification

ABSTRACT

Direct-to-PCR methodology adds samples directly to PCR tubes offering gains in efficiency and sensitivity. The approach has been applied to a variety of biological sources including blood, saliva, tissue, hair and nail. We added various preservative solutions to a range of biological samples to leech DNA into solution, whilst preserving at room temperature. Tubes containing ‘free DNA’ then followed automated workflows for amplification and capillary electrophoresis. Routine FASS-automated workflows (including DNA extraction and quantification) were compared with published direct-to-PCR methodology and automated amplification of an aliquot of preservative solution. Applying preservative solutions to ~30-year-old blood stains stored at room temperature resulted in recovery of a larger quantity of DNA and more alleles (using PowerPlex 21) when compared with routine automated typing. Trials were extended to blood, saliva, hair and nail, mimicking ante-mortem samples collected in a disaster victim identification effort. Despite slightly lower allelic recovery, the faster processing times, lower costs and storage potential offers advantages for the processing of ante-mortem samples.

KEYWORDS:

• Disaster victim identification (DVI)
• direct-to-PCR
• preservatives
• automated processing
• ante-mortem (AM)

Acknowledgements

Thermo Fisher Scientific for supplying an Identifiler Plus kit for testing.

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

This work was supported by an Australian Government Research Training Program Scholarship, and sponsored by FASS.Funding sources had no involvement in study design, collection, analysis, interpretation or writing of the report.