1. Dried blood spots of pooled samples for RHD gene screening in blood donors of mixed ancestry.
- Author
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Silva-Malta MC, Araujo NC, Vieira OV, Schmidt LC, Gonçalves Pde C, and Martins ML
- Subjects
- Female, Humans, Male, Blood Donors, Blood Grouping and Crossmatching methods, Donor Selection methods, Dried Blood Spot Testing methods, Rh-Hr Blood-Group System genetics
- Abstract
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., (© 2015 British Blood Transfusion Society.)
- Published
- 2015
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