Direct Amplification of Blood, Saliva, and Semen Subsamples Using Copan MicroFLOQ® Direct Swabs
Bibliographic record
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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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.001 | 0.002 |
| Meta-epidemiology (narrow) | 0.001 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.001 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.001 | 0.000 |
| Open science | 0.000 | 0.001 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.002 | 0.002 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".