DO/ART4 gene sequencing in sub-Saharan cohorts and African migrants: useful data describing the diversity and spreading of rare variants.

Background: Due to the unavailability of immunological reagents, the Dombrock blood group is insufficiently explored in African populations and can be a source of alloimmunization. A large study including pygmoid and nonpygmoid ethnic groups from East, Central, and West continental Africa, together with African migrants like Comorians, Afro-Caribbean from Martinique, and Maroons from French Guiana would be helpful to increase transfusion safety.

Study Design And Methods: Using genomic DNA extracted from blood samples collected from 336 nonpygmoid and 51 pygmoid Africans as well as 268 samples of African descent, DO coding regions were PCR-amplified and sequenced.

Sequencing of the ART4 gene in sub-Saharan cohorts reveals ethnic differences and two new DO alleles: DO*B-Ile5Thr and DO*B-Trp266Arg.

Background: Given the high heterogeneity of sub-Saharan populations especially between nonpygmoids and pygmoids, differences are expected during investigation of the DO/ART4 gene.

Study Design And Methods: Using genomic DNA extracted from blood samples collected from 77 Tswa pygmoids and 39 Teke and seven San nonpygmoids, DO coding regions were amplified and sequenced. A tetra-primer amplification refractory mutation system-polymerase chain reaction method was developed to specifically detect the DO*B-SH-Gln149Lys variant.

Synonymous nucleotide polymorphisms influence Dombrock blood group protein expression in K562 cells.

To gain further insight into ART4 (DO) gene alleles (DO*A, DO*JO1, DO*A-WL, DO*DOYA, DO*B, DO*B-WL, DO*B-SH-Q149K, DO*B-(WL)-I175N, DO*HY1, DO*HY2, DO*DOMR) and evaluate the impact of synonymous nucleotide polymorphisms on protein expression and mRNA accumulation of DO*A-HA, DO*A-SH and DO*B-SH alleles, human erythroleukaemic K562 cells were transducted with variant DO-lentiviral particles and analysed by flow cytometry and quantitative reverse transcription polymerase chain reaction. Monoclonal antibody (MoAb) detection of DO*A-HA and DO*JO1 transductants was lower than DO*A transductants, while detection of DO*A-SH, DO*A-WL and DO*DOYA transductants was higher. Variant DO*B alleles, i.

Characterization of novel RHD alleles: relationship between phenotype, genotype, and trimeric architecture.

Background: RH1 is one of the most clinically important blood group antigens in the field of transfusion and prevention of fetomaternal incompatibilities. New variant RHD alleles are regularly identified and their characterization is essential to ensuring patient safety.

Study Design And Methods: Blood samples with uncertain RhD phenotypes not resolved by our first-line SNaPshot assay were sequenced for all 10 RHD exons.

Dombrock genotyping in a native Congolese cohort reveals two novel alleles.

Background: Since variant alleles in the Dombrock (DO) blood group system are common in Africans, DNA typing of DO alleles in an uninvestigated Congolese Teke ethnic group was performed.

Study Design And Methods: DO exons were polymerase chain reaction amplified, using genomic DNA extracted from blood samples, and sequenced. Membrane expression in K562 cells transduced with DO-cDNAs using lentiviral vectors was studied by flow cytometry.

Using an EGFPmeter to evaluate the lentiviral vector production: tricks and traps.

An efficient viral vector containing supernatant is the first key issue to address for gene therapy or basic research projects relying on viral gene transfer. When present in a lentiviral vector backbone as a reporter gene, the expression of the enhanced green fluorescent protein (EGFP) shows strong interests to assess the production of lentiviral vector supernatants. The immediate nondestructive visual analysis of EGFP expression helps to predict the quality of the viral vector supernatant while it is produced.

Molecular analysis of inactive and active RHD alleles in native Congolese cohorts.

Background: In Africa, RHD alleles have not been fully characterized. The purpose of this study was to identify inactive and active RHD alleles at the molecular level in Congolese cohorts.

Study Design And Methods: Blood samples were collected from people living in central Congo populated by Teke ethnic group.