Claims for Patent: 9,637,788

Summary for Patent: 9,637,788

Title:	Discrimination of blood type variants
Abstract:	The present invention provides a method for detecting the presence or absence of, or for discriminating between, blood type variants, including RHD*r\'s, RHD*DIIIa, RHD*DIVa-2, RHCE*cs.sup.s and RHCE*ce733G. The method comprises genotyping a sample obtained from a human subject at one or more positions in intron 7 of the RHD gene and/or in intron 7 of the RHCE gene. The invention also provides products, in particular, probes, primers and kits for use in the method of the invention.
Inventor(s):	Ochoa; Jorge (Derio, ES), Lopez; Monica (Derio, ES), Escorza; Sergio (Derio, ES), Tejedor; Diego (Derio, ES), Martinez; Antonio (Derio, ES), Simon; Laureano (Derio, ES)
Assignee:	Progenika Biopharma, S.A. (Derio, ES)
Application Number:	14/126,819
Patent Claims:	1. A method for determining the presence or absence of, or for discriminating between, blood type alleles, which method comprises: providing a DNA-containing sample obtained from a human subject; amplifying a first segment of intron 7 of the RHD locus by allele-specific polymerase chain reaction (PCR) using a first primer comprising the nucleotide sequence of SEQ ID NO: 9 and a second primer comprising the nucleotide sequence of SEQ ID NO: 10, thereby genotyping the sample at at least the TIC polymorphic site at IVS7+1887 and at the C/A polymorphic site at IVS7+2276; amplifying a second segment of intron 7 of the RHD locus by allele-specific PCR using a first primer comprising the nucleotide sequence of SEQ ID NO: 12 and a second primer comprising the nucleotide sequence of SEQ ID NO: 13; probing said amplified second segment by allele-specific hybridization using an oligonucleotide probe having the nucleotide sequence of SEQ ID NO: 70, thereby genotyping the sample at at least the C/T polymorphic site at IVS7+3349; and predicting the blood type allele of the human subject using the genotype of the sample at said polymorphic positions at IVS7+1887, IVS7+2276 and IVS7+3349. 2. The method according to claim 1, wherein said blood type alleles are alleles that comprise an RHD/RHCE hybrid exon 3. 3. The method according to claim 1, wherein said blood type alleles are selected from the group consisting of: RHD*r's; RHD*DIIIa; RHD*DIII_FN; RHD*DIVa-2; and RHCE*ce.sup.S. 4. The method according to claim 1, wherein said method further comprises genotyping the sample at one or more polymorphic positions in intron 7 of the RHD gene locus, said one or more polymorphic positions being selected from: IVS7+1869, IVS7+1880, IVS7+1886, IVS7+2282, IVS7+3257, IVS7+3258, IVS7+3259, IVS7+3260, IVS7+3489, IVS7+3521, IVS7+3523, and IVS7+3525. 5. The method according to claim 1, wherein the method further comprises determining whether the sample contains an RHD-RHCE hybrid exon 3. 6. The method according to claim 5, wherein the method comprises genotyping the sample at one or more polymorphic positions selected from: IVS2-26, IVS2-13, IVS2-8 in RHD intron 2; IVS3+64, IVS3+69 in RHD intron 3; and position 410 in RHD exon 3. 7. The method according to claim 1, wherein the method comprises genotyping not more than 50, 40, 30, 25, 20, 15, or not more than 10, single nucleotide polymorphic positions in the RHD gene locus and/or the RHCE gene locus. 8. The method according to claim 1, wherein the method further comprises predicting an RHD phenotype and/or an RHCE phenotype for the subject based on the genotype of the sample. 9. The method according to claim 1, wherein the method further comprises determining whether the sample contains an RHD-RHCE hybrid exon 3, and wherein DNA obtained from said sample is amplified using primers that are selective for RHD/RHCE hybrid exon 3 and wherein said primers that are selective for RHD/RHCE hybrid exon 3 are selected from: a primer comprising the nucleotide sequence of SEQ ID NO: 5 and a primer comprising the nucleotide sequence of SEQ ID NO: 6. 10. The method according to claim 9, wherein said method further comprises genotyping the sample at a polymorphic position in exon 3 of the RHD locus, and wherein the method comprises an allele-specific hybridisation of DNA extracted from the sample, or an amplicon derived from DNA extracted from the sample, and wherein the allele-specific hybridisation comprises contacting said extracted DNA or said amplicon with a probe selected from: a probe comprising the sequence of SEQ ID NO: 7 and a probe comprising the sequence of SEQ ID NO: 8. 11. The method according to claim 1, further comprising genotyping the sample at one or more additional positions selected from the group consisting of IVS7+1869, IVS7+1880, IVS7+1886, IVS7+2276, IVS7+2282, and IVS7+3489. 12. The method according to claim 1, wherein the sample is determined to comprise C at position IVS7+1887, A at position IVS7+2276 and T at position IVS7+3349 and wherein the subject is predicted to have at least one RHD*r'.sup.S blood type allele. 13. The method according to claim 1, wherein the sample is determined to comprise T at position IVS7+1887, A at position IVS7+2276 and T at position IVS7+3349 and wherein the subject is predicted to have at least one RHD*DIIIa blood type allele. 14. The method according to claim 1, wherein the sample is determined to comprise T at position IVS7+1887, C at position IVS7+2276 and T at position IVS7+3349 and wherein the subject is predicted to have at least one RHD*DIII_FN blood type allele. 15. The method according to claim 1, wherein the sample is determined to comprise T at position IVS7+1887, C at position IVS7+2276 and C at position IVS7+3349 and wherein the subject is predicted to have at least one RHD*DIVa-2 blood type allele.

Details for Patent 9,637,788

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132	06/04/1986		2031-06-17
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132			2031-06-17
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	Injection	103132			2031-06-17