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Computer-assisted means for assessing lifestyle risk factors

a lifestyle risk factor and computer-aided technology, applied in the field of methods of assessing disease susceptibility, can solve the problems of increased or decreased carcinogen activation, loss of regulatory mechanisms that govern normal cell behaviour, and formation of tumours with full metastatic (or invasive) potential

Inactive Publication Date: 2006-08-10
GILL GARRISON ROSALYNN +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0030] As such sites would be able to be updated and new hyperlinks added to the report after the report is initially delivered to the client, the information and advice would be able to be updated at any time, thereby allowing the client to access up-to-date yet personalised health and dietary advice over a prolonged period, without the need for requesting another report.

Problems solved by technology

Human tumours result from a series of mutational events, leading to the loss of the regulatory mechanisms that govern normal cell behaviour and ultimately resulting in the formation of a tumour with full metastatic (or invasive) potential (Smith, 1995).
Unfortunately, the metabolic pathways do not always lead to detoxification of the toxin.
Variable levels of expression of these enzymes could result in increased or decreased carcinogen activation.
However, defining the precise roles of specific dietary factors in the development of cancer is difficult due to the multitude of variables involved (Perera, 2000).
Further complications arise due to differences in methodology, control populations, types of carcinogens, and amounts of exposure to carcinogens.
Low intake can lead to micronutrient deficiency, which has been shown to cause DNA damage in a way that mimics radiation damage by causing single and double-stranded breaks, oxidative lesions or both.
Thus, diet can be a direct supply of genotoxic compounds or may cause chronic irritation or inflammation (Giovannucci, 1999).
Such advice can therefore only be very general and cannot, by its very nature, take account of the particular genotype of an individual.
Moreover, in recent years, there has been much media publicity of research findings on links between particular foods, drugs etc and medical conditions, often causing health scares.
As the factors that contribute to disease susceptibility, for example cancer, or cardiovascular disease susceptibility vary between populations and between individuals of populations, it is often impossible for an individuals to derive useful advice appropriate to his or her particular circumstances from such reports.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

Preparation of DNA Sample

[0154] DNA is prepared from a buccal cell sample on a brush using a Qiagen QIAamp kit according to the manufacturer's instructions (Qiagen, Crawley, UK). Briefly, the brush is cut in half and one half stored at room temperature in a sealed tube in case retesting is required. The other half of the brush is placed in a microcentrifuge tube. 400 μl PBS is added and the brush allowed to rehydrate for 45 minutes at room temperature. Quiagen lysis buffer and Proteinase K is then added, the contents are mixed, and allowed to incubate at 56 C for 15 minutes to lyse the cells. Ethanol is added and the lysate transferred to a QIAamp spin column from which DNA is eluted after several washings.

example 2

Quantification of DNA

[0155] In order to check that sufficient DNA has been isolated, a quantification step is carried out using the PicoGreen dsDNA Quantification kit (Molecular Probes, Eugene, Oreg., USA).

[0156] Briefly, client DNA samples are prepared by transferring a 10 μl aliquot into a microcentrifuge tube with 90 μl TE. 100 μl of the working PicoGreen dsDNA quantification reagent is added, mixed well, and transferred into a black 96 well plate with flat well bottoms. The plate is then incubated for 5 minutes in the dark before a fluorescent reading is taken. The quantity of DNA present in the clients' samples is determined by extrapolating from a calibration plot prepared using DNA standards.

[0157] A quantity of DNA in the range of 5-50 ng total is used in the subsequent PCR step. Remaining client DNA sample is stored at −20° C. for retesting if required.

example 3

Taqman® Assay to Identify the MTHFR A1298C Polymorphism

[0158] The modified reaction mixture contains Taq polymerase (1.25 units / μl), optimised PCR buffer, dNTP (200 μM each), 2 mM MgCl2 and primer pairs SEQ ID NO: 160 and 161 and polymorphism probe SEQ ID NO: 200.

[0159] The reaction mixture is initially incubated for 10 minutes at 50° C., then 5 minutes at 95° C., followed by 40 cycles of 1 minute of annealing at between 55° C. and 60° C. and 30 seconds of denaturation at 95° C. Both during the cycles and at the end of the run, fluorescence of the released reporter molecules of the probe is measured by an integral CCD detection system of the AB7700 thermocycler. The presence of a fluorescent signal which increases in magnitude through the course of the run indicates a positive result.

[0160] The assay is then repeated with the same primer pair and wt probe SEQ ID NO: 199. If the sample is homozygous for the polymorphism, no fluorescence signal is seen with the wt probe. However, i...

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Abstract

The present invention relates to methods of assessing disease susceptibility associated with dietary and lifestyle risk factors. The invention provides for analysis of alleles at loci of genes associated with lifestyle risk factors, and the disease susceptibility profile of an individual is determined by reference to datasets which further match the risk factor with lifestyle recommendations in order to produce a personalized lifestyle advice plan.

Description

[0001] This application is a divisional of Ser. No. 09 / 771,933, filed Jan. 30, 2001 (allowed), the entire contents of which is incorporated herein by reference.FIELD OF THE INVENTION [0002] The present invention relates to methods of assessing disease susceptibility. In particular, it relates to methods of assessing disease susceptibility associated with dietary and lifestyle risk factors. BACKGROUND TO THE INVENTION [0003] Cancer is a disease influenced primarily by external factors. Up to 80% of human cancers arise from exposure to environmental agents. The majority of cancer is believed to be preventable because exposure to these external factors should be manageable (Giovannucci, 1999; Perera, 2000). [0004] Human tumours result from a series of mutational events, leading to the loss of the regulatory mechanisms that govern normal cell behaviour and ultimately resulting in the formation of a tumour with full metastatic (or invasive) potential (Smith, 1995). All higher organisms h...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): G06Q10/00G06F19/00C12N15/09C12Q1/68G06F17/30G16B25/10G16B40/10G16H10/60
CPCG06F19/20G06F19/24G06Q50/22G06Q50/24G06F19/3431G16H10/60G16H50/30G16B25/00G16B40/00G16H10/40G16H20/70G16B40/10G16B25/10G09B5/02G09B19/0092
Inventor GILL-GARRISON, ROSALYNNMARTIN, CHRISTOPHERSANCHEZ-FELIX, MANUEL
Owner GILL GARRISON ROSALYNN
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