Determination of KIR haplotypes associated with disease

• US 9,914,970 B2
• Filed: 02/03/2015
• Issued: 03/13/2018
• Est. Priority Date: 09/22/2009
• Status: Active Grant

• Current Assignee: Chldren'S Hospital & Research Center At Oakland, Illumina Incorporated, Roche Molecular Systems (Roche Holding AG)
• Original Assignee: Children's Hospital & Research Center At Oakland, Conexio Genomics Pty. Ltd. (Illumina Incorporated), Roche Molecular Systems (Roche Holding AG)
• Inventors: Ladner, Martha, Trachtenberg, Elizabeth, Bentley, Lloyd Gordon, Goodridge, Damian, Erlich, Henry A.
• Primary Examiner(s): Woolwine, Samuel C
• Application Number:
  US14/613,290
• Publication Number:
  US 20150184243A1
• Time in Patent Office: 1,134 Days
• Field of Search: None
• CPC Class Codes: C12Q 1/6881 for tissue or cell typing, e.g. human leukocyte antigen [HLA] probes
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers
• CPC Class Codes: C12Q 2600/16 Primer sets for multiplex assays
• CPC Class Codes: C12Q 2600/172 Haplotypes
• CPC Class Codes: Y02A 90/10 Information and communication technologies [ICT] supporting adaptation to climate change, e.g. for weather forecasting or climate simulation

Abstract

Disclosed is a method of determining KIR genotypes for one or more individuals in parallel, the method comprising: for each individual, amplifying the polymorphic exon sequences of the KIR genes, pooling the KIR amplicons, performing emulsion PCR followed by pyrosequencing in parallel to determine all the amplicon sequences present in the individual to determine which KIR alleles are present in the individual.

Claims

1. A kit for obtaining KIR amplicons to determine sequences of the same exon from multiple KIR genes in one or more individuals in parallel, comprising:

a forward primer comprising an adapter region, an individual identification tag and a KIR-hybridizing region capable of hybridizing to and supporting amplification of sequences of the same exon from multiple KIR genes;

a reverse primer that comprises an adapter region, an individual identification tag selected from Table 3, and a KIR-hybridizing region capable of hybridizing to and supporting amplification of more than one KIR gene.

2. The kit of claim 1, further comprising one or more populations of beads having a primer attached, said primer capable of hybridizing to the adapter regions in said forward and reverse primers.

3. The kit of claim 1, wherein the KIR-hybridizing regions of the forward and reverse primers are selected from Table 2.