Molecular RH blood group typing of serologically D-/CE+ donors: the use of a polymerase chain reaction-sequence-specific primer test kit with pooled samples

Donatella Londero, Mauro Fiorino, Valeria Miotti, Vincenzo de Angelis
2019 Immunohematology  
The known presence of RHD blood group alleles in apparently D-individuals who are positive for C or E antigens leads to an appropriate investigation for the RHD gene on the red blood cells (RBCs) of D-blood donors, thus preventing their RBCs from immunizing D-recipients. Ready-to-use polymerase chain reaction-sequence-specific primer (PCR-SSP) typing kits are available and allow single-sample results. The need to perform this testing on a large number of donors affiliated with the Transfusion
more » ... partment of Udine (Northern Italy) led to the use of molecular genetic RH blood group typing with PCR-SSP test kits and DNA samples mixed in pools. From a population of 35,000 blood donors screened for D antigen by serologic typing, a total of 235 samples, distributed in pools of 5 DNA samples, were investigated. Positive results were reevaluated by opening the pools and retesting single samples. Validation of DNA-pool typing with commercial kits was done. Among 235 genotyped samples, 12 were found to be PCR positive (5.1%), exhibiting DEL genotype and RHD-CE-D hybrid alleles. Our data demonstrate that the use of a PCR-SSP commercial test kit with pooled samples is a helpful and valid method to correctly detect RHD alleles. As a consequence, we reclassified our donors as carriers of potentially immunogenic alleles. Immunohematology 2011;27:25-28.
doi:10.21307/immunohematology-2019-171 fatcat:ddtjlyrwprcrvlwds7pfv4hg4i