Molecular genetic blood group typing by the use of PCR-SSP technique.
BACKGROUND
DNA-based methods are useful for enhancing immunohematology typings. Ready-to-use Conformité Européenne (CE)-marked test kits based on polymerase chain reaction with sequence-specific priming (PCR-SSP) have been developed, which enable the examination of weak, unexpected, or unclear serologic findings. DEVELOPMENT AND VALIDATION: Primers were designed according to established mutation databases. Proficiency testing for CE marking was performed in accordance with Directive 98/79EC of the European Parliament and of the Council of October 27, 1998 on in vitro diagnostic medical devices using pretyped in-house and external samples. INTENDED USE: BAGene PCR-SSP kits are in vitro diagnostic devices. Genotyping of ABO and RHD/RHCE as well as HPA and KEL, JK, and FY specificities has to be performed after the conclusion of the serologic determination.


CONCLUSIONS
Ready-to-use PCR-SSP typing kits allow the determination of common, rare, or weak alleles of the ABO blood group, Rhesus, and Kell/Kidd/Duffy systems as well as alleles of the human platelet antigens.


RESULTS
The investigations showed clear-cut results in accordance with serology or molecular genetic pretyping.


CONCLUSIONS
PCR-SSP is a helpful supplementary technique for resolving most of the common problems caused by discrepant or doubtful serologic results, and it is an easy-to-handle robust method. Questionable cases in donor, recipient, and patient typing can be examined with acceptable cost.
Prager
Martina
M
Department of Production Molecular Genetics, BAG--BiologischeAnalysensystemGmbH, Lich, Germany. prager.martina@bag-germany.com
D000017:ABO Blood-Group System / Q000032:analysis; D001788:Blood Grouping and Crossmatching / Q000379:methods*; D017931:DNA Primers / Q000138:chemical synthesis; D006580:Genetic Carrier Screening; D005838:Genotype; D006801:Humans; D007626:Kell Blood-Group System / Q000032:analysis; D007667:Kidd Blood-Group System / Q000032:analysis; D008954:Models, Biological; D008967:Molecular Biology / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D018807:Polymorphism, Single-Stranded Conformational; D012204:Rh-Hr Blood-Group System / Q000032:analysis
D000017:ABO Blood-Group System; D017931:DNA Primers; D007626:Kell Blood-Group System; D007667:Kidd Blood-Group System; D012204:Rh-Hr Blood-Group System