Robotic High Thoroughput Multiplex PCR Single-Nucleotide Polymorphism Genotyping of Apheresis Platelet Donors.
Bibliographic record
Abstract
Abstract Abstract 2106 Poster Board II-83 Background Phenotyping blood donors for human platelet antigens (HPAs) can be limited by the availability of antibodies and methodologies. Robotic high throughput multiplex polymerase chain reaction (PCR) single-nucleotide polymorphism (SNP) technology provides the opportunity to perform mass genotyping for HPAs to screen apheresis donors who are negative for antigens implicated in alloimmune thrombocytopenia. This technology needs to be validated by standard polymerase chain reaction-SSP to establish a repository of platelet donors to allow for timely and adequate platelet support for patients requiring HPA-matched platelets. The objective of this study is to validate the use of high throughput SNP technology. Methods This is a prospective cohort study of 750 regular apheresis donors. Platelet apheresis donors were identified from Canadian Blood Services' Toronto Centre and genotyped for HPA 1-5 and 15 using high throughput SNP (SNPStream®) technology. HPAs were genotyped using both the sense and antisense DNA strands of each polymorphism to ensure maximum specificity. The genotypes identified by SNP were confirmed by standard methods thus all donors found to be negative for antigens implicated in alloimmune thrombocytopenia were retested using manual (sequence specific SSP-PCR at Canadian Blood Services' Platelet Immunology Laboratory and at the Toronto Centre. Results Of the 750 donors that were screened, 130 donors were found to be negative for antigens implicated in alloimmune thrombocytopenia based on SNP technology. The table illustrates genotyping results using SNP technology and SSP-PCR confirmation to date. In some instances, donors were homozygous for two low frequency HPA genotypes: 2 donors were homozygous for both HPA-1b/b and -2b/b, 6 donors for HPA-1b/b and -3b/b, 1 donor for HPA-2b/b and -3b/b, 1 donor for HPA-1b/b and -5b/b, 10 donors for HPA-1b/b and -15b/b, 4 donors for HPA-5b/b and -15b/b and 1 donor for HPA-2b/b and 15-b/b. HPA System Number of Genotypes Identified by SNP Technology Discrepant results Identified by SSP HPA-1 b/b 15 2 (13%) HPA-2 b/b 8 3 (27%) HPA-3 b/b 25 1 (4%) HPA-5 b/b 1 0 HPA-15 b/b 9 1 (11%) Conclusions: Robotic high throughput multiplex PCR SNP is potentially useful for mass genotyping of apheresis platelet donors and can identify both low frequency antigen-negative donors and combinations of these genotypes. This technology needs to be further developed. Disclosures: No relevant conflicts of interest to declare.
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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 | 0.001 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.001 | 0.001 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.001 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.005 | 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".