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Abstract

Direct amplification workflows generate DNA profiles from small amounts of biological material by omitting the steps of DNA extraction and quantitation, and thus have the potential to expedite DNA profile generation in forensic laboratories. The study herein explored how microFLOQ^® direct swabs perform in both routine forensic DNA testing (i.e., polymerase chain reaction capillary electrophoresis, or PCR-CE) and using a novel direct amplification massively parallel sequencing (MPS) protocol. First, the quality of DNA profiles generated via direct amplification of subsamples of blood, saliva, and semen were evaluated, as well as the quantity of DNA recovered during this subsampling. Cotton swabs and 4N6FLOQSwabs^® were used to collect liquid blood, saliva, and semen and were subsequently subsampled (i.e., onto a new medium) using single collection, nylon tip microFLOQ^® direct swabs. Subsamples were processed immediately after fluid deposition (i.e., wet subsamples) and after 1 week of drying (i.e., dried subsamples), and DNA profiles were generated with PCR-CE. Subsamples of fresh bodily fluids collected from cotton swabs yielded a greater number of complete short tandem repeat (STR) profiles when compared with subsamples of dried bodily fluids, while dried blood yielded only incomplete STR profiles via direct amplification. Subsamples of fresh semen taken from 4N6FLOQSwabs^® yielded complete but, in many instances, oversaturated STR profiles, suggesting that very little sample is required from a neat semen stain. Direct quantitation results for fresh and dried blood, saliva, and semen were found to not inform the quality or completeness of the resultant STR profiles. In a separate experiment, 5% and 10% dilutions of saliva and blood, as well as touch DNA samples, were collected from nonabsorbent surfaces using microFLOQ^® direct swabs, where DNA profiles were generated using a custom MPS protocol. In sum, direct amplification of microFLOQ^® direct subsamples taken from cotton swabs, 4N6FLOQSwabs^®, and nonabsorbent surfaces produced high-quality STR profiles suitable for forensic interpretation.

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Direct Amplification of Blood,Saliva,and Semen Subsamples Using Copan MicroFLOQ® Direct Swabs

Abstract

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References

Supplementary Material