Method for the Simultaneous Determination of Blood Group and Platelet Antigen Genotypes

Abstract

RBC and platelet (Plt) alloimmunization requires antigen-matched blood to avoid adverse transfusion reactions. Some blood collection facilities use unregulated Abs to reduce the cost of mass screening, and later confirm the phenotype with government approved reagents. Alternatively, RBC and Plt antigens can be screened by virtue of their associated single nucleotide polymorphisms (SNPs). We developed a multiplex PCR-oligonucleotide extension assay using the GenomeLab SNPStream platform to genotype blood for a plurality of blood group antigen-associated SNPs, including but not limited to: RhD (2), RhC/c, RhE/e, S/s, K/k, Kp^a/b, Fya/b, FYO, Jk^a/b, Di^a/b, and HPA-1a/b.

Description

TECHNICAL FIELD[0001]This invention relates to an ultra high throughput (UHT) multiplex PCR genotyping method. More specifically, the present invention relates to an automated method of determining a plurality of blood group and platelet antigen, preferably human platelet antigen (HPA), genotypes simultaneously from a single sample through the detection of single nucleotide polymorphisms (SNPs) for various blood group and platelet antigens.BACKGROUND OF THE INVENTION[0002]At present, there are 29 blood group systems and 6 HPA systems recognized by the International Society of Blood Transfusion (ISBT), wherein, with a few exceptions, a blood group ‘system’ may be defined by a single gene at a given locus of the human genome (Daniels, G. L. et al. Vox Sang 2003; 84:244; Metcalfe P. et al., Vox Sang. 2003; 85:240). Most people know their ABO and Rh blood group. However, the ABO and Rh blood group systems expressed on red cells simply represent antigens from only two of the 29 blood gro...

AI Technical Summary

Benefits of technology

[0013]The present invention provides a method of detecting the presence or absence of nucleotides relating to various SNPs for the determination of a specific genotype and accordingly the inferred phenotype. More specifically, the present invention allows for the detection of the presence or absence of two nucleotides of a plurality of different SNPs, and more preferably of the 12 SNPs in a preferred embodiment of the present invention.

Problems solved by technology

The gold standard in the industry is to ‘phenotype’ blood for the presence of specific blood group and platelet antigens using government regulated antisera (antibodies) performed by single-test methods or by an automated platform, which is a cost ineffective method for a blood collection facility that routinely performs tests on a high volume basis.

Essentially, the current tests which employ government-approved reagents in a manual, single-test driven method are a very cost ineffective method for a blood collection facility that is often required to perform such tests on a high volume basis.

However, since much of the blood is sent to hospitals within 24-48 hours after collection, manual blood group phenotyping cannot meet the short turn-around time required to provide the end user with the information required before blood must be shipped.

The prior art does not provide the use of multiple DNA sequences as primer pairs that work simultaneously on a single sample.

Moreover, the prior art does not employ novel DNA sequences to detect blood group SNPs in an automated high-throughput fashion.

The deficiencies of these test systems are the use of a single PCR reaction for each nucleotide of a given nucleotide of each SNP, and the pooling of samples prior to the detection phase and manual post-analyte data analysis.

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