First investigation of RH gene polymorphism in patients with sickle cell disease and associated blood donors in Cameroon, Central Africa

Abstract

Background - Although genetic polymorphism of the RH blood group system is well known in sub-Saharan Africa, national/regional specificities still remain to be described precisely. For the first time in Cameroon, Central Africa, and in order to better characterize the molecular basis driving RH phenotype variability, as well as to identify the main antigens that may be potentially responsible for alloimmunization, we sought 1/ to study the RH genes in a cohort of 109 patients with sickle cell disease; 2/ to analyse similarly the corresponding donors whose red blood cells (RBCs) were transfused to the patients (108 donors in 98 patients); 3/ to predict RH phenotype on the basis of the molecular data and compare with serologic testing; and 4/ to identify retrospectively patients at risk for alloimmunization.

Materials and methods - In order to generate an exhaustive dataset, the RH genes of all patient and donor samples were systematically investigated by 1/ quantitative multiplex PCR of short fluorescent fragments (QMPSF) for characterization of RHD gene zygosity and potential structural variants (SVs), and 2/ Sanger sequencing for identification of single nucleotide variants (SNVs). Following molecular analysis, genotypes and RH phenotype were deduced and predicted from reference databases, respectively.

Results- In a total of 217 Cameroonian individuals, as many as 24 and up to 22 variant alleles were identified in the RHD and RHCE genes, respectively, in addition to the reference alleles. Interestingly, 65 patients with SCD (66.3%) were assumed to be exposed to one or more undesirable RH antigen(s) with various clinical relevance.

Discussion - Beyond the comprehensive report of the nature and distribution of RH variant alleles in a subset of Cameroonian patients treated by transfusion therapy, this work highlights the requirement for an extensive review of the current practice, including routine serologic typing procedures, in a near future.