How do we identify RHD variants using a practical molecular approach?. Issue 4 (28th February 2014)

• Record Type: Journal Article
• Authors: Arnoni, Carine Prisco; Latini, Flavia Roche Moreira; Muniz, Janaína Guilhem; Gazito, Diana; Person, Rosangela de Medeiros; de Paula Vendrame, Tatiane Aparecida; Barreto, José Augusto; Castilho, Lilian
• Is Part Of: Transfusion. Volume 54:Issue 4(2014)
• Page Start: 962
• Page End: 969
• Publication Date: 2014-02-28
• Subjects: Hematology -- Periodicals; Blood -- Transfusion -- Periodicals; Blood Group Antigens -- Periodicals; Blood Preservation -- Periodicals; Blood Transfusion -- Periodicals; 615
• DOI: 10.1111/trf.12557
• Languages: English
• ISSNs: 0041-1132

Abstract

<abstract abstract-type="main"> <title> <x xml:space="preserve">Abstract</x> </title> <p>Serologic resolution of Rh discrepancies due to partial D or weak D phenotypes is a frequent problem encountered during routine typing that can be solved by <italic>RHD</italic> genotyping because it provides better characterization of these variants. The objective of the current study was to develop algorithms for identification of D variants in multiethnic populations based on a logic sequence of molecular tests using a large number of atypical RhD specimens. Thus, a total of 360 blood samples with atypical D antigen expression were analyzed. A previously published multiplex polymerase chain reaction (PCR) procedure was performed and depending on multiplex PCR analysis, the associated <italic>RHCE</italic> allele, and D variant frequency in our population, an algorithm was developed composed of six flow charts using specific PCR–restriction fragment length polymorphism and/or specific exon sequencing. This strategy allowed the identification of 22 different variants with few assays and a much reduced cost. This study describes a simple and practical algorithm that we use to determine <italic>RHD</italic> genotypes in samples with unknown <italic>RHD</italic>. This strategy is relatively easy to implement and the algorithm can be adapted to populations with various ethnic backgrounds after an initial assessment of the type and frequency of D variants. Essentially, we demonstrate that sequencing of all <italic>RHD</italic> exons is not necessary for the identification of the majority of known D variants.</p> </abstract>