Discriminating braf mutations

Patent metadata

Field	Value
Publication number	US-2020115760-A1
Application number	US-201916577510-A
Country	US
Kind code	A1
Filing date	Sep 20, 2019
Priority date	Feb 24, 2014
Publication date	Apr 16, 2020
Grant date	—

Abstract

Provided herein are methods for detecting and discriminating BRAF V600 mutations. Also provided herein are methods for diagnosis, prognosis, management, and treatment decisions of BRAF V600 mutation-related diseases or conditions.

First claim

1 - 19 . (canceled)

20 . A kit for detecting a V600E or V600K mutation in the BRAF gene in a sample DNA from an individual, said kit comprising:
(a) a forward primer and a reverse primer, wherein the forward primer and the reverse primer are configured to amplify DNA sequences containing a thymidine (T) to adenosine (A) mutation at position 1799 of the BRAF gene, wherein the primers amplify both a V600E (1799T>A) mutation and a V600K (1798-1799GT>AA) mutation in the BRAF gene and do not amplify the wild type BRAF gene, and (b) a restriction enzyme, wherein the restriction enzyme digests an amplicon comprising the BRAF V600E mutation (1799T>A) and does not digest an amplicon comprising the BRAF V600K mutation (1798_1799GT>AA).

21 . The kit of claim 20 , wherein the restriction enzyme recognizes a 6-base motif that matches the sequence of the BRAF gene immediately upstream of the thymidine (T) to adenosine (A) mutation at position 1799 of the BRAF gene.

22 . The kit of claim 20 , wherein the restriction enzyme is SfcI.

23 . The kit of claim 20 , wherein the forward and/or reverse primer are labeled with a detectable moiety.

24 . The kit of claim 23 , wherein the detectable moiety is a fluorophore.

25 . The kit of claim 20 , wherein the sample DNA is extracted from the plasma of the individual.

26 . The kit of claim 20 , wherein the individual is a melanoma patient.

Assignees

• Quest Diagnostics Invest Llc

Inventors

• Donahue Amber C
• Peng Yen-Lin

Classifications

• C12Q2600/118

  Prognosis of disease development · CPC title

• C12Q2600/156

  Polymorphic or mutational markers · CPC title

• C12Q1/6886 Primary

  for cancer (immunoassay for cancer G01N33/575) · CPC title

• C12Q2600/106

  Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism · CPC title