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Diagnostic method

a cancer and prognosis technology, applied in the field of cancer diagnosis, prognosis or monitoring, can solve the problems of methods plagued by the number of false positives generated, and achieve the effect of assessing the effectiveness of treatment in the individual and predicting the susceptibility of individuals to cancer

Inactive Publication Date: 2014-09-18
THE CHINESE UNIVERSITY OF HONG KONG
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Benefits of technology

The present invention provides a method for detecting or monitoring cancer using a biological sample from an individual. The method involves obtaining DNA from the sample, digesting it with methylation-sensitive restriction enzymes, and quantifying or detecting a DNA sequence of interest that contains at least two recognition sites for the enzyme. The DNA sequence can be a target sequence that is methylated in cancer patients but not in normal individuals. The method can be used to predict the susceptibility to cancer, assess the stage of cancer, predict the likelihood of overall survival or recurrence, and assess the effectiveness of treatment. The invention also provides probes, primers, and kits for use in the method.

Problems solved by technology

However, as noted by the authors Yegnasubramanian et al, Nucleic Acids Research, Vol. 34, No. 3, 2006 e19, such methods are plagued by the number of false-positives that are generated.

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[0156]Sixty-three hepatocellular carcinoma (HCC) patients, as well as two groups of controls, were recruited. The first control group consisted of 15 healthy control subjects without chronic hepatitis B infection. The second control group consisted of chronic hepatitis B carriers. For each HCC patient, one sex-and age-matched chronic hepatitis B carrier was recruited as a control. Chronic hepatitis B carriers had increased risk of developing HCC and, thus, would be the target group for HCC screening. Four milliliters of venous blood were collected from each study subject into EDTA-containing tubes. Blood samples were centrifuged at 1,600 g for 10 min and the supernatant was re-centrifuged at 16,000 g for 10 min. DNA was then extracted from 800 μL of plasma using the QIAamp mini kit (Qiagen, Hilden, Germany) and eluted with 50 μL of H2O. Thirty-five microliters of plasma DNA were digested with 100 U of BstU I enzyme, in 1× digestion buffer at 60° C. for 16 hours.

[0157]The concentrati...

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Abstract

The present invention concerns a method for the detection or monitoring of cancer using a biological sample selected from blood, plasma, serum, saliva, urine from an individual, said method comprising:(a) obtaining DNA from the said biological sample;(b) digesting the DNA sample with one or more methylation-sensitive restriction enzymes;(c) quantifying or detecting a DNA sequence of interest after step (b), wherein the target sequence of interest contains at least two methylation-sensitive restriction enzyme recognition sites; and(d) comparing the level of the DNA sequence from the individual to a normal standard, to detect, prognosticate or monitor cancer.

Description

CROSS REFERENCE TO RELATED PATENT APPLICATION [0001]This application claims the benefit of the filing date of U.S. provisional patent application 60 / 847,499, filed Sep. 27, 2006, the disclosure of which is herein incorporated by reference.FIELD OF THE INVENTION [0002]The present invention relates to methods for diagnosis, prognosis or monitoring of cancer in an individual, in particular using the differential methylation patterns in genes associated with cancers.BACKGROUND TO THE INVENTION [0003]It is well known that many tumor suppressor genes are methylated in tumor cells. As such, the use of methylation markers has been suggested for the detection or monitoring of cancer in patients. A number of different methods have been proposed for detection of these methylated sequences.[0004]Methylation specific PCR (MSP) is the most commonly used method for detecting methylated or unmethylated DNA. MSP involves the step of bisulfite conversion. Sodium bisulfite is used to deaminate cytosin...

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/118C12Q2600/154
Inventor LO, YUK MING DENNISCHAN, KWAN CHEE ALLENDING, CHUNMING
Owner THE CHINESE UNIVERSITY OF HONG KONG
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