RESUMEN
BACKGROUND AND OBJECTIVES: Next generation sequencing (NGS) has promising applications in transfusion medicine. Exome sequencing (ES) is increasingly used in the clinical setting, and blood group interpretation is an additional value that could be extracted from existing data sets. We provide the first release of an open-source software tailored for this purpose and describe its validation with three blood group systems. MATERIALS AND METHODS: The DTM-Tools algorithm was designed and used to analyse 1018 ES NGS files from the ClinSeq® cohort. Predictions were correlated with serology for 5 antigens in a subset of 108 blood samples. Discrepancies were investigated with alternative phenotyping and genotyping methods, including a long-read NGS platform. RESULTS: Of 116 genomic variants queried, those corresponding to 18 known KEL, FY and JK alleles were identified in this cohort. 596 additional exonic variants were identified KEL, ACKR1 and SLC14A1, including 58 predicted frameshifts. Software predictions were validated by serology in 108 participants; one case in the FY blood group and three cases in the JK blood group were discrepant. Investigation revealed that these discrepancies resulted from (1) clerical error, (2) serologic failure to detect weak antigenic expression and (3) a frameshift variant absent in blood group databases. CONCLUSION: DTM-Tools can be employed for rapid Kell, Duffy and Kidd blood group antigen prediction from existing ES data sets; for discrepancies detected in the validation data set, software predictions proved accurate. DTM-Tools is open-source and in continuous development.
Asunto(s)
Alelos , Antígenos de Grupos Sanguíneos/análisis , Antígenos de Grupos Sanguíneos/genética , Secuenciación del Exoma/métodos , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Programas Informáticos , Sistema del Grupo Sanguíneo Duffy/genética , Variación Genética , Técnicas de Genotipaje , Humanos , Glicoproteínas de Membrana/genética , Proteínas de Transporte de Membrana/genética , Metaloendopeptidasas/genética , Receptores de Superficie Celular/genética , Transportadores de UreaRESUMEN
CONCLUSIONS: Isoagglutinins in the plasma of apheresis platelets are a concern. High titer anti-A and anti-B can cause severe hemolytic transfusion reactions. Our facility is testing donor plasma using the gel method to identify isoagglutinin titers exceeding 250. Platelet additive solution (PAS), recently introduced as a collec-tion and storage solution, replaces approximately 65 percent of the plasma in a platelet component. We intended to confirm the effect of PAS on the isoagglutinin titers. We compared the isoagglutinin titers in donor plasma from EDTA-anticoagulated whole blood (without PAS) with the plasma in apheresis platelet components with PAS. Titers were determined in a buffered gel matrix test using serial twofold dilution steps. Among 100 donors tested, 26 plasma samples exceeded a threshold titer of 250; 25 were group O and only one was group B. When samples from these 26 platelet components with PAS were tested, only one group O donor exceeded the threshold titer. Samples from plasma components with PAS consistently showed a 50 percent decrease in titer compared with the donors' plasma samples. In conclusion, nearly half of the group O donors tested exceeded a titer of 250. Only one apheresis platelet component with PAS exceeded this clinically applied threshold-a 96 percent decrease compared with the number of donor plasma samples without PAS. The implementation of PAS in apheresis platelet components prompted us to revise our component screening process, which then minimized component manipulation of out-of-group platelet transfusions.