Application of the Luminex Platform for Multiplexed Genotyping of Non-Deletional Alpha Thalassemia Mutations.
Bibliographic record
Abstract
Abstract Objectives: Certain point mutations in the alpha globin gene may lead to unstable hemoglobins such as Hb Constant-Spring and Hb Quong-Sze. These alpha chain variants, when inherited with deletional α-thalassemia mutations cause more severe forms of HbH disease. To evaluate the utility of the Luminex xMAP platform for rapid screening of non-deletional α-thalassemia mutations, we developed a multiplexed genotyping assay to detect six mutations in the α2-globin gene. that are clinically significant or, when coinherited with α-thalassemia-1, result in HbH disease. Methods: A panel of positive controls with known heterozygosity for Hb Constant-Spring, Hb Quong-Sze, Hb J-Toronto, Hb Hasharon, Hb G-Philadelphia and Hb G-Norfolk, was obtained from anonymized whole blood or newborn dried blood spots. The α2 gene was amplified using nested PCR and single base chain extension (SBCE) was carried out using the PCR products, capture-tagged primers, thermosequenase, and one of four biotinylated ddNTPs. Biotinylated amplicons were hybridized to fluorescent microspheres containing allele-specific capture probes and analyzed by flow cytometry on the Luminex100. Results: Median flourescent intensity (MFI) values uniformly discriminated between wild type and mutant alleles in three independent experiments. Mean background-subtracted MFIs ranged from 255 to 1000 for all positive alleles and genotypes were correctly identified in all of the samples tested. Conclusions: As HbH disease is becoming more prevalent in the U.S., early detection of this disorder using accurate, affordable diagnostic methods will permit timely delivery of health care and prevention of severe clinical manifestations of disease. As the severity of many of these disorders has not been well-characterized, an understanding the phenotypic expression of these hemoglobin disorders is essential to providing adequate counseling and treatment options.
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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 | 0.003 | 0.001 |
| Meta-epidemiology (narrow) | 0.001 | 0.001 |
| Meta-epidemiology (broad) | 0.001 | 0.000 |
| Bibliometrics | 0.001 | 0.001 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.002 | 0.001 |
| Open science | 0.001 | 0.001 |
| Research integrity | 0.001 | 0.001 |
| Insufficient payload (model declined to judge) | 0.003 | 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".