US 11,453,908 B2Granted

Methods and kits for typing KIR2DL alleles

AssigneeMemorial Sloan Kettering Cancer Center

InventorsKatharine Hsu, Jean-Benoit Le Luduec

Life of this patent

Estimating the term…

Abstract

Disclosed herein are methods and compositions such as primers, primer pairs, and kits for typing KIR2DL1, KIR2DL2, Sand KIR2DL3 alleles. The compositions and methods disclosed herein are useful in the selection of most appropriate donors for HCT.

Claims11

Dependent claims

A method for determining the allelic group of KIR2DL1 alleles in a subject, comprising:

(a) performing PCR reactions in a genomic DNA sample obtained from the subject with
(i) a first reverse primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
4 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 4011 of SEQ ID NO:
66 and a first forward primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
3 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 3680 of SEQ ID NO:
66 that are configured to generate a first amplification product, if present;
(ii) a second reverse primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
6 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 5820 of SEQ ID NO:
66 and a second forward primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
5 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 5499 of SEQ ID NO:
66 that are configured to generate a second amplification product, if present;
(iii) a third reverse primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
8 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 13609 of SEQ ID NO:
66 and a third forward primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
7 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 13420 of SEQ ID NO:
66 that are configured to generate a third amplification product, if present;
(iv) a fourth reverse primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
10 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 5735 of SEQ ID NO:
66 and a fourth forward primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
9 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 5499 of SEQ ID NO:
66 that are configured to generate a fourth amplification product, if present;
and
(v) a fifth reverse primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
12 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 4011 of SEQ ID NO:
66 and a fifth forward primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
11 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 3790 of SEQ ID NO:
66 that are configured to generate a fifth amplification product, if present;
and
(vi) a sixth reverse primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
14 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 5761 of SEQ ID NO:
66 and a sixth forward primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
13 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 5616 of SEQ ID NO:
66 that are configured to generate a sixth amplification product, if present;
and
(b) determining one or more allelic groups of KIR2DL1 alleles present in the subject based on detection of the first, second, third, fourth, fifth, and sixth amplification products from the PCR reactions, wherein the one or more allelic groups of KIR2DL1 alleles are selected from among group 1 (*003), group 2 (*006), group 3 (*012), group 4 (*004), group 5 (*010) and group 6 (*002).

32 more elements

Dependencies are inferred from the claim text.

Facts

Application: 16/338,667Filed 28 Sep 2017 · us provisional application
Priority: 1 Oct 2016
Published: US 2019/0226017 A125 Jul 2019
Granted: 27 Sep 2022
Term ends: Not estimatedThe source records don’t support an estimate.

Classification10 codes

C12Q 1/6858
Nucleic acid amplification reactions: allele-specific amplification
C12Q 1/6806
Involving nucleic acids: preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
C12Q 1/6827
Hybridisation assays: for detection of mutation or polymorphism
C12Q 1/6853
Nucleic acid amplification reactions: using modified primers or templates

Family

Application62/403,099Us provisional application · filed 1 Oct 2016
Application62/403,131Us provisional application · filed 1 Oct 2016
US 2019/0226017 A1Pre-grant publication · published 25 Jul 2019
US 11,453,908 B2This patent

From the US filing history. Foreign family members aren’t listed.

Timeline

Citations

Open elsewhere

patentgrep

Loading patent…

Abstract

A method for determining the allelic group of KIR2DL1 alleles in a subject, comprising:

(a) performing PCR reactions in a genomic DNA sample obtained from the subject with
(i) a first reverse primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
4 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 4011 of SEQ ID NO:
66 and a first forward primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
3 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 3680 of SEQ ID NO:
66 that are configured to generate a first amplification product, if present;
(ii) a second reverse primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
6 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 5820 of SEQ ID NO:
66 and a second forward primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
5 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 5499 of SEQ ID NO:
66 that are configured to generate a second amplification product, if present;
(iii) a third reverse primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
8 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 13609 of SEQ ID NO:
66 and a third forward primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
7 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 13420 of SEQ ID NO:
66 that are configured to generate a third amplification product, if present;
(iv) a fourth reverse primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
10 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 5735 of SEQ ID NO:
66 and a fourth forward primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
9 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 5499 of SEQ ID NO:
66 that are configured to generate a fourth amplification product, if present;
and
(v) a fifth reverse primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
12 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 4011 of SEQ ID NO:
66 and a fifth forward primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
11 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 3790 of SEQ ID NO:
66 that are configured to generate a fifth amplification product, if present;
and
(vi) a sixth reverse primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
14 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 5761 of SEQ ID NO:
66 and a sixth forward primer that comprises at least 10 nucleotides that are identical to SEQ ID NO:
13 and binds specifically to a region of KIR2DL1 alleles comprising nucleotide 5616 of SEQ ID NO:
66 that are configured to generate a sixth amplification product, if present;
and
(b) determining one or more allelic groups of KIR2DL1 alleles present in the subject based on detection of the first, second, third, fourth, fifth, and sixth amplification products from the PCR reactions, wherein the one or more allelic groups of KIR2DL1 alleles are selected from among group 1 (*003), group 2 (*006), group 3 (*012), group 4 (*004), group 5 (*010) and group 6 (*002).

32 more elements

Facts

Application

16/338,667Filed 28 Sep 2017 · us provisional application

Priority

1 Oct 2016

Published

US 2019/0226017 A125 Jul 2019

Granted

27 Sep 2022

Term ends

Not estimatedThe source records don’t support an estimate.

Classification10 codes

C12Q 1/6858
Nucleic acid amplification reactions: allele-specific amplification
C12Q 1/6806
Involving nucleic acids: preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
C12Q 1/6827
Hybridisation assays: for detection of mutation or polymorphism
C12Q 1/6853
Nucleic acid amplification reactions: using modified primers or templates