Concordance of two polymerase chain reaction–based blood group genotyping platforms for patients with sickle cell disease

Abstract

In recent years, polymerase chain reaction-based genotyping platforms, which provide a predicted phenotype, have increased in both patient and high-throughput donor testing, especially in situations where serologic methods or reagents are limited. This study looks at the concordance rate between two platforms commercially available in the United States when used for testing samples from patients with sickle cell disease (SCD), a group particularly vulnerable to alloimmunization. DNA extracted from samples from 138 patients with SCD was tested by human erythrocyte antigen (HEA) BeadChip (Immucor, Norcross, GA) and by ID CORE XT (Progenika-Grifols, Barcelona, Spain). Predicted phenotype results were compared, and a concordance rate was calculated. Discrepancies were resolved by Sanger sequencing. All testing was done under an institutional review board-approved protocol. A concordance rate of 99.9 percent was obtained. Sanger sequencing was performed on four samples with discrepancies in the Rh blood group system. Three samples had a similar allelic variant detected by ID CORE XT. Two of the three discrepant samples were correctly identified as V+w, VS- by ID CORE XT but not by HEA BeadChip. The third sample, predicted to have a phenotype of V+, VS+ by sequencing, was called correctly by HEA BeadChip but not by ID CORE XT, which had predicted V+w, VS-. The fourth discrepancy was identified in a sample that ID CORE XT accurately identified as RHCE*ce[712G] and predicted a partial c phenotype. This result was confirmed by Sanger sequencing, whereas HEA BeadChip found no variants and predicted a c+ phenotype. The high concordance rate of the two methods, along with the known limitations of serology, warrant further discussion regarding the practice of serologic confirmation of extended phenotypes. Clinical significance of the identified discrepancies remains to be determined.