DETERMINATION OF THE GENOTYPE UNDERLYING THE S-s-U- PHENOTYPE OF THE MNSs BLOOD GROUP SYSTEM

A polynucleotide capable of at least partially or fully hybridizing to a hybrid nucleotide sequence representative of a deletional mutation of the GYPB gene wherein the hybrid nucleotide sequence representative of a deletional mutation of the GYPB gene has a sequence according to SEQ ID NO 1 or SEQ ID NO 2, or a complementary sequence thereto, under stringent conditions.

A method for testing for the presence of a polynucleotide comprising a hybrid nucleotide sequence representative of a deletional mutation of the GYPB gene, wherein the hybrid nucleotide sequence representative of a deletional mutation of the GYPB gene has a sequence according to SEQ ID NO 1 or SEQ ID NO 2, in a sample comprising genomic DNA or an amplification product thereof, comprising contacting the genomic DNA or an amplification product thereof of the sample with one or more polynucleotide probes capable of at least partially or fully hybridizing to the hybrid nucleotide sequence representative of a deletional mutation of the GYPB gene, under stringent conditions, and detecting any hybridization of said one or more polynucleotide probes to the hybrid nucleotide sequence representative of a deletional mutation of the GYPB gene.

A primer pair for use in an amplification reaction, comprising a forward and a reverse primer, wherein the forward primer is capable of hybridizing to a region upstream of a polynucleotide comprising a hybrid nucleotide sequence representative of a deletional mutation of the GYPB gene, wherein the hybrid nucleotide sequence representative of a deletional mutation of the GYPB gene has a sequence according to SEQ ID NO 1 or SEQ ID NO 2, under stringent conditions, and wherein the reverse primer is capable of hybridizing to a region downstream of a polynucleotide comprising a hybrid nucleotide sequence representative of a deletional mutation of the GYPB gene, wherein the hybrid nucleotide sequence representative of a deletional mutation of the GYPB gene has a sequence according to SEQ ID NO 1 or SEQ ID NO 2, under stringent conditions.

A kit for testing for the presence of a polynucleotide comprising a hybrid nucleotide sequence representative of a deletional mutation of the GYPB gene, wherein the hybrid nucleotide sequence representative of a deletional mutation of the GYPB gene has a sequence according to SEQ ID NO 1 or SEQ ID NO 2, in a sample comprising genomic DNA or an amplification product thereof, comprising one or more polynucleotide probes capable of at least partially or fully hybridizing to said hybrid nucleotide sequence representative of a deletional mutation of the GYPB gene under stringent conditions, in separate containers.

A kit for identifying a human carrier of a double deletional mutation of the GYPB gene causative of a S-s-U− phenotype, comprising,

• a. a first container comprising a first primer pair of oppositely oriented primers, wherein the primers are capable of hybridizing to a template DNA at a region upstream of a hybrid nucleotide sequence representative of a deletional mutation of the GYPB gene according to SEQ ID NO1 and at a region downstream of said hybrid nucleotide sequence, respectively, under stringent conditions;
• b. a second container comprising a second primer pair of oppositely oriented primers, wherein the primers are capable of hybridizing to a template DNA at a region upstream of a hybrid nucleotide sequence representative of a deletional mutation of the GYPB gene according to SEQ ID NO2 and at a region downstream of said hybrid nucleotide sequence, respectively, under stringent conditions;
• c. a third container comprising a third primer pair of oppositely oriented control primers capable of hybridizing to a undeleted, wildtype nucleotide sequence not represented in either of the hybrid nucleotide sequences, such as for example the intact GYPB gene.

A method for determining whether a patient in need of a blood transfusion is to be transfused with blood from a donor of a S-s-U− phenotype, comprising the step of testing a sample of the patient genomic DNA, or an amplification product thereof, for the presence of a polynucleotide comprising a hybrid nucleotide sequence representative of a deletional mutation of the GYPB gene according to SEQ ID NO1 or SEQ ID NO2.

A method for determining whether blood of a donor may be used for transfusion of a patient of a S-s-U− phenotype in need thereof, comprising the step of testing a sample of the donor genomic DNA, or an amplification product thereof, for the presence of a polynucleotide comprising a hybrid nucleotide sequence representative of a deletional mutation of the GYPB gene according to SEQ ID NO1 or SEQ ID NO2.

A method of assessing the risk of a mother of a S-s-U− phenotype of conceiving or carrying a foetus of a U+ phenotype or the risk of a mother having an anti-U titer of conceiving or carrying a foetus at risk of developing haemolytic disease of the new-born, comprising testing a sample of the genomic DNA, or an amplification product thereof, of the father of the foetus or of the foetus itself for the presence of a polynucleotide comprising a hybrid nucleotide sequence representative of a deletional mutation of the GYPB gene according to SEQ ID NO1 or SEQ ID NO2.

A method for determining whether a human is a carrier of a double deletional mutation of the GYPB gene causative of a S-s-U− phenotype, comprising the steps of a. obtaining the nucleotide sequences of at least the GYP locus on both maternal and paternal chromosomes of said individual in a machine-readable format, b. determining if the obtained sequences comprise a hybrid nucleotide sequence representative of a deletional mutation of the GYPB gene according to SEQ ID NO1 or SEQ ID NO2 on either maternal or parental chromosome, or both, and c. providing the corresponding genotype.

A method of determining whether a human is a carrier of at least one deletional mutation of the GYPB gene or of a double deletional mutation of the GYPB gene causative of a S-s-U− phenotype, comprising testing the genomic DNA, or an amplification product thereof, of said human for the presence of a polynucleotide comprising a hybrid nucleotide sequence representative of a deletional mutation of the GYPB gene according to SEQ ID NO1 or SEQ ID NO2.

A kit for identifying a human carrier of at least one deletional mutation of the GYPB gene, comprising

• a. a first container comprising a first primer pair of oppositely oriented primers, wherein the primers are capable of hybridizing to the template DNA at a region upstream of a hybrid nucleotide sequence representative of a deletional mutation of the GYPB gene according to SEQ ID NO1 and at a region downstream of said hybrid nucleotide sequence, respectively, under stringent conditions;
• b. a second container comprising a second primer pair of oppositely oriented primers, wherein the primers are capable of hybridizing to the template DNA at a region upstream of a hybrid nucleotide sequence representative of a deletional mutation of the GYPB gene according to SEQ ID NO2 and at a region downstream of said hybrid nucleotide sequence, respectively, under stringent conditions.

A method for determining whether a human is a carrier of a double deletional mutation of the GYPB gene causative of a S-s-U− phenotype, comprising the steps of a. obtaining the nucleotide sequences of the whole genome of said individual in a machine-readable format, b. determining if the obtained sequences comprise a hybrid nucleotide sequence representative of a deletional mutation of the GYPB gene according to SEQ ID NO1 or SEQ ID NO2 on either maternal or parental chromosome, or both, and c. providing the corresponding genotype.