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Methods of detecting colorectal cancer

Inactive Publication Date: 2012-06-28
EXACT SCI CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0022]Also described herein is a method of sample processing, prior to carrying out a method of the invention, comprising removing a portion of a collected faecal sample and adding the removed portion of the sample to a buffer which prevents denaturation or degradation of blood proteins found in the sample.

Problems solved by technology

Procedures such as digital rectal examination (DRE); colonoscopy or sigmoidoscopy are highly invasive, painful and can cause a great deal of patient discomfort.
While sensitive and specific, the procedure is invasive, costly, has limited availability and includes certain risks such as induction of infection and perforation of the bowel.
Immunochemical tests (FIT or iFOBT) that use anti-hemoglobin antibodies specific for human blood in extracts from stool do not require dietary restrictions; however, they are more complicated and more expensive than peroxidase-based tests.
In addition, human hemoglobin in fecal samples degrades with time, resulting in a loss of antigenicity which can produce false negative results.
While this test is more sensitive than fecal occult blood testing, it is not as sensitive as colonoscopy and will miss about half of cancers in an average risk group of people without symptoms.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

Sample Collection and Processing

[0147]A standardized multicenter screening trial (The Netherlands) was initiated in 2006. In this trial, non symptomatic subjects aged 50 or above are screened with colonoscopy, FOBT and real-time MSP using DNA from stool and blood. In addition, prospectively collected stool samples from multiple centers (Germany and The Netherlands) were used. In these trials, symptomatic patients, attending a Gastroenterology clinic and ultimately diagnosed with CRC, provided a stool sample for use in real-time MSP. From the ongoing trials 111 stool samples were available for the present study. 3 main categories of stool samples were used: 38 samples with no suspicious findings, 10 adenomas and 63 samples from patients covering all stages of CRC.

[0148]After defecation in a special bucket, an iFOBT samples was taken (Eiken device) and the patients subsequently added 250 ml of stool homogenization buffer (Amresco, Solon, Ohio, USA) to the sample. Samples were shipped ...

example 2

FIT and Methylation Testing on Partial Stool Samples

[0174]In Example 1 homogenates of whole stool sample material were investigated. In this example, the inventor evaluated whether sufficient DNA material could be subtracted from partial stool samples allowing subsequent methylation analysis.

Sample Collection and Nucleic Acid Isolation

[0175]82 historical stool samples, collected between 2003 and 2008, were available for the present study. The samples were frozen down at the time of collection without addition of stabilizing buffer and stored at −20° C. until further use.

[0176]In this study only a portion of the stool sample (≈1 g) was investigated for further methylation testing. First, historical samples were thawed at the storage site and partial samples were taken (≈1 g) using special designed stool tubes with integrated measuring spoons (Sarstedt). Each sample was then weighted and 2 volumes of stabilizing buffer (Diagn Mol Pathol; Volume 14, Number 3, September 2005) were added...

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Abstract

A method of detecting a predisposition to, or the incidence of colorectal cancer in a faecal sample comprises, in a first step (a), detecting the presence of blood in the faecal sample, wherein detection of the presence of blood is indicative of a predisposition to, or the incidence of colorectal cancer. The method additionally comprises, in second step (b), detecting an epi-genetic modification in the DNA contained within the faecal sample, wherein detection of the epigenetic modification is indicative of a predisposition to, or the incidence of colorectal cancer. Based upon a positive result obtained in either (a) or (b) or in both (a) and (b) a predisposition to, or the incidence of colorectal cancer is detected. Related methods and kits involve detecting an epigenetic modification in a number of specific genes.

Description

FIELD OF THE INVENTION[0001]The present invention is concerned with the diagnosis, staging and treatment of disease, in particular cancer and more specifically colorectal cancer. The invention relates to methods and kits for diagnosing colorectal cancer based upon detecting epigenetic modifications, typically in specific genes. The methods and kits may also permit the detection of blood in a fecal sample, with the combined tests proving particularly advantageous.BACKGROUND OF THE INVENTION[0002]Colorectal cancer (CRC) is a leading cause of cancer-related deaths worldwide, and is the second leading cause of cancer-related deaths in the United States. A patient's prognosis is good if the cancer is caught early, when the site of the cancer is confined to its site of origin. However, the cure rates fall once the cancer has spread. Most colon cancers arise from conventional adenomatous polyps (conventional adenoma-to-carcinoma sequence), while some colon cancers appear to arise from the ...

Claims

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Application Information

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IPC IPC(8): A61K33/36A61P35/00C12Q1/68
CPCC12Q1/6886C12Q2600/106G01N2800/52C12Q2600/158G01N2800/50C12Q2600/154A61P35/00G01N33/57446G01N2333/805
Inventor JOOST, LOUWAGIE
Owner EXACT SCI CORP
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