Dried blood spots of pooled samples for <scp><i>RHD</i></scp> gene screening in blood donors of mixed ancestry

Silva-Malta, Maria Clara Fernandes da; Araujo, N. C. Fidélis; Vieira, Otavio Vinicius Neves; Schmidt, Luciana Cayres; Gonçalves, Poliane de Cássia; Martins, Marina Lobato

doi:10.1111/tme.12235

Cited by 2 publications

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“…The use of specific tests for the detection or confirmation of the presence of RHDψ is essential for the inference of the RhD phenotypes in the Brazilian population because the presence of this variant has already been described in this population (Rodrigues et al, ; Cruz et al, ). In Minas Gerais, situated in Southeast Brazil, a recent study demonstrated that 3·74% of the serologically D‐, C‐ and E ‐negative blood donors had the RHD gene, and 3·02% of them carried non‐functional RHDψ (Silva‐Malta et al, ). In the present study, 1·34% of the blood donors randomly selected according to the RhD phenotype ( n = 289) tested positive for RHDψ .…”

Section: Discussionmentioning

confidence: 99%

Molecular analysis of the RHD pseudogene by duplex real‐time polymerase chain reaction

Silva-Malta

Santos

Gonçalves

et al. 2019

Transfusion Medicine

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Objectives:In this study, we aimed to present a strategy for the detection of the RHD pseudogene (RHD ) based on a real-time polymerase chain reaction (PCR) assay. Background:The D-negative phenotype is associated with many genetic alterations. In populations with African ancestry, this phenotype commonly results from the silent variant RHD . The evaluation of RHD is essential for correct inference of the RhD phenotype in order to avoid false-positive results. Methods:We utilised a new method for the simultaneous detection of RHD and a fragment from exon 5 of the wild-type RHD gene based on duplex real-time PCR assay. Results:The PCR assay allowed specific detection of RHD . There was complete agreement between the results generated by the new test and the results generated by molecular analysis performed using end-point PCR methods previously described. Conclusions:The assay developed is easy to execute and presents the potential for routine use at blood banks and other associated facilities where it is desired to determine the presence of RHD .

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Section: Discussionmentioning

confidence: 99%

Molecular analysis of the RHD pseudogene by duplex real‐time polymerase chain reaction

Silva-Malta

Santos

Gonçalves

et al. 2019

Transfusion Medicine

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“…The use of DBS on filter paper as the source of DNA facilitates the transport and storage of samples, eliminating the need for refrigeration, and has been successfully used in many areas, including blood group analysis . The genotyping of pooled samples allows cost reduction, which is desirable in rare blood donor programmes as it is necessary to screen thousands of individuals to find a rare donor .…”

Section: Discussionmentioning

confidence: 99%

The screening of rare blood donors in a highly admixed population: A new approach for Holley and Diego genotyping and impact of genomic and self‐reported ancestry

Muniz

Silva

Ferraz

et al. 2019

Transfusion Medicine

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Objectives: The present study aimed to develop strategies for genotyping DO*HY (Dombrock system) and DI*A/DI*B (Diego system) alleles and to evaluate the impact of genomic and self-declared ancestry on rare donor screening in admixed populations.Background: The antigens Hy and Di b demonstrate clinical importance. The lack of antisera for the serological evaluation of these antigens makes it necessary to develop molecular methods. In addition, considering that some rare red blood cell phenotypes present differences in frequency between ethnic groups, it is important to assess the applicability of self-declared ancestry in the search for rare donors in admixed populations.Methods: DO*HY and DI*A/DI*B genotyping based on real-time polymerase chain reaction (PCR) was standardised. A total of 457 blood donors clustered by selfdefined skin colour/race categories were genotyped. Furthermore, individual genomic ancestry was used in the analyses. Results:The assays developed are reproducible and provide satisfactory results even at low concentrations of DNA, which make them useful in situations where the DNA is scarce, such as dried blood spots on filter paper, or when screening for pooled samples. No significant difference was observed in the frequencies of the DI*A, DI*B and DO*HY, comparing the self-declared White (branco) donors with those who are Black (preto) and Brown (pardo). Conclusion:Real-time PCR, especially using pooled samples, is a promising strategy to screen rare blood donors. Although both self-reported race/colour and some blood group phenotypes are associated with ancestry, the results point to a greater complexity in the application of self-declared race/colour in the screening of rare donors in admixed populations. K E Y W O R D S ancestry, Diego, Dombrock, rare blood donor Amanda A. Muniz and Adão R. da Silva contributed equally to this study.

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

Abstract: The present method is a promising strategy to detect the RHD gene among presumed RhD-negative blood donors, particularly for populations with African ancestry.

Cited by 2 publications

References 20 publications

Molecular analysis of the RHD pseudogene by duplex real‐time polymerase chain reaction

Molecular analysis of the RHD pseudogene by duplex real‐time polymerase chain reaction

The screening of rare blood donors in a highly admixed population: A new approach for Holley and Diego genotyping and impact of genomic and self‐reported ancestry

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