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US 5,516,634 AGranted
Molecular basis of the human platelet bra/brb alloantigen system and applications thereof
OwnerPeter J. Newman, Sentot S. Santosoinventors
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OwnerPeter J. Newman, Sentot S. Santosoinventors
Estimating the term…
Based on the discovery of the amino acid and nucleotide differences which distinguish the Br.sup.a and Br.sup.b allelic forms of the membrane glycoprotein GPIa, and which comprise the diallelic Br platelet alloantigen system, compositions and methods are provided for determining the Br genotype and phenotype of individuals. Also provided, on the basis of this discovery, are compositions and methods for treating disorders associated with Br alloantigen incompatibility, such as the bleeding disorders post-transfusion purpura, post-transfusion platelet refractoriness, and neonatal alloimmune thrombocytopenic purpura. The two allelic forms of GPIa differ by a single amino acid. The Br.sup.a allelic form has Lys at position 505 in the sequence of the mature GPIa. The Br.sup.b allelic form has Glu at the same position. This amino acid difference is due to a single change, from A for the Br.sup.a allele to G for the Br.sup.b allele, in the GPIa gene.
An oligonucleotide which has a sequence selected from the group consisting of (a) a subsequence of 8-50 nucleotides of SEQ ID NO:1, which subsequence includes the A at position 1600 of SEQ ID NO:1;
A test kit for Br alloantigen typing, comprising:
A method of Br alloantigen typing comprising the steps of (A) obtaining, by a target nucleic acid amplification process applied to mRNA from human platelets or human genomic DNA, an assayable quantity of amplified nucleic acid with a sequence that is a subsequence, or the full complement of a subsequence, of the mRNA or genomic DNA that encodes a GPIa subunit of a Br alloantigen, said subsequence including the nucleotide at the position in the mRNA or genomic DNA corresponding to the position of 5'-nucleotide of the codon for the amino acid at position 505 of the mature GPIa subunit;
In combination in an aqueous solution, a pair of oligonucleotide primers for the amplification by the PCR process of a segment of platelet mRNA which encodes a GPIa or a segment of genomic DNA for GPIa, said oligonucleotide primers being selected from the group consisting of DNAs with the sequences of SEQ ID NO:2-SEQ ID NO:5, SEQ ID NO:11 and SEQ ID NO:12.
A method of amplifying platelet mRNA which comprises amplification by the PCR method employing as primers a pair of oligonucleotide primers for the amplification by the PCR process of a segment of platelet mRNA which encodes a GPIa subunit, said oligonucleotide primers being selected from the group consisting of DNAs with the sequences of SEQ ID NO:2-SEQ ID NO:5.
Dependencies are inferred from the claim text.