Detection of 13q14 chromosomal alterations

• US 7,479,370 B2
• Filed: 09/08/2004
• Issued: 01/20/2009
• Est. Priority Date: 09/08/2003
• Status: Expired due to Fees

• Current Assignee: Health Research Incorporated
• Original Assignee: Health Research Incorporated
• Inventors: Coignet, Lionel J.
• Primary Examiner(s): Sitton; Jehanne S
• Application Number:
  US10/937,514
• Publication Number:
  US 20050079531A1
• Time in Patent Office: 1,595 Days
• Field of Search: None
• US Class Current: 435/6.11
• CPC Class Codes: C12Q 1/6841 In situ hybridisation
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 2600/112 Disease subtyping, staging or classification

Abstract

The present invention relates to methods for detection of chromosomal alterations which are associated with the presence of various leukemias and lymphomas. The method comprises the steps of obtaining a biological sample comprising lymphocytes from an individual and assaying the sample to detect chromosomal deletions in the regions of chromosome 13 that corresponds to the region of chromosome 13 present in the RP11-147H23 or RP11-327P2.

Claims

1. A method of diagnosing an individual human as likely of having lymphoblastic leukemia (ALL) comprising the steps of:

a) obtaining a sample of bone marrow or blood comprising lymphocytes from the individual, and

b) detecting by fluorescence in situ hybridization (FISH) whether part of the 13q14 region of chromosome 13 present in bacterial artificial chromosome (BAC) RP11-147H23 is deleted in the lymphocytes, wherein the FISH analysis is performed using a test probe and a control probe, wherein the test probe is BAC RP11-147H23, and wherein the control probe is complementary to a sequence of chromosome 13 outside the region of chromosome 13 present in BAC RP11-147H23;

wherein, in the FISH analysis, two pairs of co-localized signals from the test probe and control probe in the lymphocytes is indicative of the absence of a deletion, and wherein a split signal from the test probe and one pair of co-localized signals is indicative of the presence of a deletion;

wherein a deletion detected in step b) is indicative of a likelihood of the individual as having ALL.

2. The method of claim 1, wherein the test probe and control probe are each labeled with a distinct fluorochrome-conjugated nucleotide.

3. The method of claim 1, wherein the control probe is either BAC RP11-215B15 or RP11-131F1.

4. A method of diagnosing a human—

individual as likely of having CLL comprising the steps of;

a) obtaining blood or bone marrow cells comprising lymphocytes from the individual;

b) isolating mRNA from lymphocytes; and

c) analyzing from b) WDFY2 mRNA expression,

wherein absence of WDFY2 mRNA expression is indicative of the individual as likely of having CLL.

5. The method of claim 4, wherein analyzing WDFY2 mRNA expression is performed using RT-PCR, wherein—

the RT-PCR is performed using a first primer consisting of the sequence of SEQ ID NO;

1 and a second primer consisting of the sequence of SEQ ID NO;

2.