Method for preparing fecal sample, fecal sample preparation solution, and feces collection kit

A preparation method and feces technology, which can be applied in the fields of biochemical equipment and methods, microbial determination/inspection, recombinant DNA technology, etc., which can solve the problems of missed detection of colon cancer and low sensitivity.

Inactive Publication Date: 2011-07-20
OLYMPUS CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in fecal occult blood testing, there is a problem that the sensitivity is as low as about 25%, and the probability of missing colon cancer is high

Method used

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  • Method for preparing fecal sample, fecal sample preparation solution, and feces collection kit
  • Method for preparing fecal sample, fecal sample preparation solution, and feces collection kit
  • Method for preparing fecal sample, fecal sample preparation solution, and feces collection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0156] Each 0.5 g of feces collected from one healthy person was put into eight 15 mL polypropylene tubes. To each stool, 10 mL of the preparation solution S described in Table 1 was added to sufficiently disperse the stool, and stool samples 1-1 to 1-8 were prepared, respectively. In addition, the pH of the preparation solution S to which 0.1M citric acid was added was adjusted to the pH shown in Table 1 with NaOH. In addition, the pH of the 60% ethanol solution to which no organic acid was added was the pH before addition to the feces.

[0157] [Table 1]

[0158] Solution for preparation

water soluble organic solvent

organic acid

pH

Stool sample 1-1

60% ethanol

0.1M citric acid

3.0

[0159] Stool sample 1-2

60% ethanol

0.1M citric acid

3.5

Stool samples 1-3

60% ethanol

0.1M citric acid

4.0

Stool samples 1-4

60% ethanol

0.1M citric acid

4.5

Stool samples 1-5 ...

Embodiment 2

[0167] As the organic acid added to the preparation solution S, adipic acid was used instead of citric acid, and the nucleic acid preservation effect at this time was investigated. The RNA recovery step and the PCR step were carried out in the same manner as in Example 1, except that the solutions described in Table 2 were used as the preparation solution S. In addition, the pH adjustment of the preparation solution S to which 0.1 M adipic acid was added was performed with NaOH. On the other hand, the pH of the preparation solution S to which 0.1 M acetic acid was added was adjusted using sodium acetate, and the pH of the preparation solution S to which 0.1 M lactic acid was added was adjusted using sodium lactate.

[0168] [Table 2]

[0169] Solution for preparation

water soluble organic solvent

organic acid

pH

Stool sample 2-1

60% ethanol

0.1M adipic acid

4.5

Stool sample 2-2

60% ethanol

0.1M adipic acid

5.0

...

reference example 1

[0173] Each 1 g of feces collected from one healthy person was collected into three 15 mL polypropylene tubes. One of the tubes was immediately frozen with liquid nitrogen to make a stool sample (1A). For the other tube, add 10 mL of 70% ethanol solution to fully disperse the feces, and then let it stand at room temperature for 1 hour to make a feces sample (1B). The remaining one tube was collected without adding a solution, etc., and quickly transferred to the extraction process, which was used as a stool sample (1C).

[0174]Then, RNA was recovered from each stool sample. Specifically, 3 mL of a phenolic mixture "Trizol" (manufactured by Invitorogen) was added to each stool sample, and they were thoroughly mixed with a homogenizer for 30 seconds or more. 3 mL of chloroform was added to the mixed solution, and after mixing well with a vortex mixer, centrifugation was performed at 12,000×g and 4° C. for 20 minutes. The supernatant (aqueous layer) obtained by the centrifuga...

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Abstract

Provided is a method for preparing a fecal sample which can stably store a nucleic acid contained in feces without the need of any complicated procedure. Also disclosed is a fecal sample preparation solution to be used in the method. Further disclosed is a feces collection kit. Still further disclosed is a method for collecting a nucleic acid from feces by using a fecal sample prepared by the above-mentioned method and analyzing the nucleic acid. Feces collected is mixed with a fecal sample preparation solution containing, as an active ingredient, a water-soluble organic solvent containing an organic acid, thereby preparing a fecal sample which can store a nucleic acid contained therein satisfactorily.

Description

technical field [0001] The present invention relates to a method for preparing a stool sample, a solution for preparing a stool sample, a kit for collecting stool, and a stool sample prepared by the preparation method for preparing a stool sample having excellent preservation properties of nucleic acids contained in the stool sample. A stool sample, a method for recovering nucleic acid from the stool sample, and a nucleic acid analysis method using the nucleic acid recovered by the nucleic acid recovery method. [0002] This application claims the priority of Japanese Patent Application No. 2008-217019 for which it applied to Japan on August 26, 2008, and uses the content here. Background technique [0003] Like Europe and the United States, the number of patients with colon cancer in Japan is also increasing rapidly year by year, and colon cancer has already occupied the top few cancer mortality rates. The reason for this is believed to be that the eating habits of the Jap...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09C12Q1/68
CPCC12Q1/6806C12Q2527/125
Inventor 谷上恭央长冈智纪
Owner OLYMPUS CORP
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