<abstract xmlns="http://www.w3.org/1999/xhtml"><p>The known presence of <italic>RHD</italic> blood group alleles in apparently D– individuals who are positive for C or E antigens leads to an appropriate investigation for the <italic>RHD</italic> 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 Department of Udine (Northern Italy) led to the use of molecular genetic <italic>RH</italic> 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 <italic>DEL</italic> genotype and <italic>RHD-CE-D</italic> 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 <italic>RHD</italic> alleles. As a consequence, we reclassified our donors as carriers of potentially immunogenic alleles. <bold>Immunohematology 2011;27:25–28.</bold></p></abstract>

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 Department 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.

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

Immunohematology

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

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...

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

Article Category: Case Report

Publicado en línea: 01 dic 2019

Páginas: 25 - 28

DOI: https://doi.org/10.21307/immunohematology-2019-171

© 2011 D. Londero, et al., published by Sciendo