Cost-efficient sequence-specific priming-polymerase chain reaction screening for blood donors with rare phenotypes.
BACKGROUND
Transfusion support for patients with irregular antibodies to red blood cell (RBC) antigens of high frequency may be hampered by lack of appropriate antigen-negative RBC units. Often, this perceived lack is due to the low number of typed donors. We developed a simple multiplex polymerase chain reaction (PCR) method to screen for donors with rare blood group phenotypes.


METHODS
A multiplex PCR with sequence-specific priming predicting Yt(a), Co(a), Lu(b), and Kp(b) antigens was developed based on a commercially available system (Extract-N-Amp, Sigma-Aldrich) that obviates the DNA purification step. PCR amplicons were analyzed by size fractionation in a 2 percent agarose gel. Samples representing rare phenotypes were identified by the lack of one of the four visible bands. Donors of blood phenotype O D- ccddee were screened.


RESULTS
Excluding the preparation of the reaction mixture and the gel, the whole procedure consisted of five pipetting steps. Hands-on time was 102 minutes for 91 donors. After optimization, interpretable results were obtained in 85 percent of samples without repetition. Among 3422 donors tested, 1 Kp(b-), 6 Co(a-), 10 Yt(a-), and 5 Lu(b-) donors were detected.


CONCLUSIONS
Multiplex PCR is a simple, versatile, and cost-efficient method for the screening for donors with rare phenotypes who may be identified by their genotype. If such donor screening is introduced on a broad basis, transfusion support for patients with anti-Co(a), anti-Yt(a), or anti-Lu(b) will be considerably improved.
Wagner
Franz F
FF
DRK Blutspendedienst NSTOB, Institut Springe, Springe, Germany. fwagner@bsd-nstob.de
Bittner
Rita
R
Petershofen
Eduard K
EK
Doescher
Andrea
A
Müller
Thomas H
TH
D001483:Base Sequence; D001782:Blood Donors; D006801:Humans; D008403:Mass Screening; D010641:Phenotype; D016133:Polymerase Chain Reaction / Q000191:economics* / Q000379:methods*; D013997:Time Factors