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

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Immunohematology

American National Red Cross

Subject: Medical Laboratory Technology

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ISSN: 0894-203X
eISSN: 1930-3955

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VOLUME 27 , ISSUE 1 (March 2011) > List of articles

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

Keywords : Rh blood group, D antigen expression, blood donors, PCR-SSP test kit

Citation Information : Immunohematology. Volume 27, Issue 1, Pages 25-28, DOI: https://doi.org/10.21307/immunohematology-2019-171

License : (Transfer of Copyright)

Published Online: 01-December-2019

ARTICLE

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.

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