Simultaneous detection of mutational status and gene copy number

Patent metadata

Field	Value
Publication number	US-2016130666-A1
Application number	US-201614997544-A
Country	US
Kind code	A1
Filing date	Jan 17, 2016
Priority date	Dec 31, 2009
Publication date	May 12, 2016
Grant date	—

Abstract

The present invention provides compositions and methods for simultaneously detecting mutational status and gene copy number. In particular, the present invention provides simultaneous measurement of gene copy number and detection of the L858R and Exon 19 del mutations in a tissue sample.

First claim

1 . A method for assessing the EGFR status of a tissue sample comprising: processing said tissue sample with reagents to produce distinguishable signals corresponding to the presence or absence of a L858R EGFR mutation, an exon 19 deletion EGFR mutation and EGFR gene amplification; and simultaneously visualizing said distinguishable signals.

2 . The method of claim 1 , wherein said tissue sample is obtained from a subject suspected of having cancer, a subject diagnosed with cancer, or a subject suffering from cancer.

3 . The method of claim 1 , wherein said processing comprises contacting said sample with antigen binding molecules specific for EGFR molecules comprising the L858R and/or exon 19 deletion mutation.

4 . The method of claim 3 , wherein said processing further comprises contacting said sample with nucleic acid probes specific for the EGFR gene.

5 . The method of claim 4 , wherein said processing further comprises contacting said antigen binding molecules specific for EGFR molecules comprising the L858R and/or exon 19 deletion mutation and said nucleic acid probe specific for the EGFR gene with reagents that produce a detectable signal corresponding to the presence or absence of said EGFR L858R and/or exon 19 deletion mutation and the presence or absence of EGFR gene amplification.

6 . The method of claim 5 , wherein said antigen binding molecules specific for EGFR molecules comprising the L858R and/or exon 19 deletion mutation and said nucleic acid probe specific for the EGFR gene comprise a signal generating moiety.

7 . The method of claim 5 , wherein said antigen binding molecules specific for EGFR molecules comprising the L858R and/or exon 19 deletion mutation and said nucleic acid probe specific for the EGFR are detected with signal generating systems.

8 . The method of claim 7 , wherein said signal generating systems comprise reagents for the differential detection of said antigen binding molecules specific for EGFR molecules comprising the L858R and/or exon 19 deletion mutation and said nucleic acid probe specific for the EGFR.

9 . The method of claim 8 , wherein said signal generating system comprises reagents for generating different colorimetric signals for each of the said antigen binding molecules specific for EGFR molecules comprising the L858R and/or exon 19 deletion mutation and said nucleic acid probe specific for the EGFR.

10 . The method of claim 9 , wherein said reagents for generating different colorimetric signals are selected from the group consisting of silver, fast red, fast blue, fast gold, DAB, AP orange, and AP blue.

11 . The method of claim 9 , wherein said reagents for generating different colorimetric signals comprise enzymatically labeled reagents.

12 . The method of claim 11 , wherein the enzyme labels of said enzymatically labeled reagents are selected from the group consisting of horseradish peroxidase, alkaline phosphatase, acid phosphatase, glucose oxidase, β-galactosidase, β-glucouronidase and β-lactamase.

13 . The method of claim 1 , further comprising: evaluating changes in EGFR gene copy number and the presence or absence of EGFR mutations.

14 . The method of claim 13 , further comprising using said evaluation to make a diagnosis of prognosis for the patient.

15 . The method of claim 13 , further comprising using said evaluation to determine a therapeutic treatment.

16 . A kit comprising: a first antigen binding molecule specific for EGFR molecules comprising a L858R mutation; a second antigen binding molecule specific for EGFR molecules comprising an exon 19 deletion mutation; a nucleic acid probe specific for EGFR; and distinguishable signal-generating reagents specific for each of said first antigen binding molecule, said second antigen binding molecule, and said nucleic acid probe.

Assignees

• Ventana Med Syst Inc

Inventors

• Singh Shalini
• Nitta Hiro
• Gaire Fabien
• Del Valle Edmundo David

Classifications

• G01N33/5759

  involving compounds localised on the membrane of tumour or cancer cells · CPC title

• G01N33/57492

  Physics · mapped topic

• C12Q2600/118

  Prognosis of disease development · CPC title

• C12Q2600/156

  Polymorphic or mutational markers · CPC title

• C12Q2600/158

  Expression markers · CPC title