Non-invasive comparative study of HLA genotyping between urinary and blood DNA using sequencing-based typing and third-generation sequencing
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Tie Cheng Sun [1] ; Cheng Yan Fan [1] ; Yu Jie Wen [1] ; Dong Mei Li [1] ; Yuan Yuan Jing [1] ; Na Liu [1] ; Jie Wang [1] ; Li Jun Wang [1] ; Xue Lian [1] ; Yan Jun Jia [1]
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[1] Beijing Red Cross Blood Center
Beijing Red Cross Blood Center
China
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- Localización: Advances in Laboratory Medicine / Avances en Medicina de Laboratorio, ISSN-e 2628-491X, Vol. 7, Nº. 1, 2026, págs. 52-58
- Idioma: inglés
- Enlaces
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Resumen
Objectives: To evaluate the feasibility of urinary DNA as a noninvasive alternative for high-resolution HLA genotyping and validate its concordance with conventional blood-based methods.
Methods: Matched urine and blood samples were collected from 11 healthy volunteers. Urinary DNA was extracted using an optimized column-based protocol, while blood DNA was processed via an automated system. High-resolution HLA typing for HLA-A, -B, -C, -DRB1, and -DQB1 loci was performed using sequencing-based typing (SBT) and thirdgeneration sequencing (TGS), with concordance rates assessed between sample types.
Results: The average concentration of urinary DNA exhibited significantly lower concentrations than blood DNA (9.74 ± 10.52 vs. 33.13 ± 26.78 ng/μL, p = 0.001) but comparable purity (OD 260/280 ratio: 1.65 ± 0.4 vs. 1.81 ± 0.13, p = 0.068).
Remarkably, both SBT and TGS achieved 100 % concordance between urine- and blood-derived genotypes across five classical HLA loci, with TGS resolving full-length HLA sequences (5′UTR–3′UTR) at ≥30× coverage.
Conclusions: Urinary DNA achieves blood-comparable accuracy in noninvasive HLA genotyping. Our optimized protocol overcomes DNA yield and purity limitations through dual-platform validation (SBT/TGS), establishing urine as a clinically viable alternative for HLA profiling, especially where blood sampling is unfeasible.
