Dried blood spots of pooled samples for RHD gene screening in blood donors of mixed ancestry

• Published in: Transfusion medicine (Oxford, England) England), 2015-10, Vol.25 (5), p.337-341
• Main Authors: Silva-Malta, M. C. F., Fidélis Araujo, N. C., Neves Vieira, O. V., Cayres Schmidt, L., de Cassia Gonçalves, P., Lobato Martins, M.
• Format: Article
• Language: English
• Subjects: Blood Donors; Blood Grouping and Crossmatching - methods; Donor Selection - methods; Dried Blood Spot Testing - methods; Female; Humans; immunohematology; Male; molecular testing; Rh blood group; Rh-Hr Blood-Group System - genetics
• ISSN: 0958-7578; 1365-3148
• DOI: 10.1111/tme.12235

SUMMARY Objectives In this study, we present a strategy for RHD gene screening based on real‐time polymerase chain reaction (PCR) using dried blood spots of pooled samples. Background Molecular analysis of blood donors may be used to detect RHD variants among the presumed D‐negative individuals. RHD genotyping using pooled samples is a strategy to test a large number of samples at a more reasonable cost. Materials and Methods RHD gene detection based on real‐time PCR using dried blood spots of pooled samples was standardised and used to evaluate 1550 Brazilian blood donors phenotyped as RhD‐negative. Positive results were re‐evaluated by retesting single samples using real‐time PCR and conventional multiplex PCR to amplify five RHD‐specific exons. PCR‐sequence‐specific primers was used to amplify RHDψ allele. Results We devised a strategy for RHD gene screening using dried blood spots of five pooled samples. Among 1550 serologically D‐negative blood donors, 58 (3·74%) had the RHD gene. The non‐functional RHDψ allele was detected in 47 samples (3·02%). Conclusion The present method is a promising strategy to detect the RHD gene among presumed RhD‐negative blood donors, particularly for populations with African ancestry.