Method for detecting disease-related marker using gastric mucosal lavage fluid

A technology of markers and gastric mucosa, which is applied in the field of detection of disease-related markers, can solve problems such as retention, inability to extract Southern blots, diagnostic possibility and diagnostic accuracy, and achieve the effect of simple collection and avoiding pollution

Inactive Publication Date: 2009-05-27
SAPPORO MEDICAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for example, in the method described in Non-Patent Document 7, for about half of the samples, it was not possible to extract a sufficient amount of DNA to perform the Southern blot method required for diagnosis. There are problems in the degree, and it is still far from the level of practical application.

Method used

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  • Method for detecting disease-related marker using gastric mucosal lavage fluid
  • Method for detecting disease-related marker using gastric mucosal lavage fluid
  • Method for detecting disease-related marker using gastric mucosal lavage fluid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] Embodiment 1: Specimen collection container

[0089] figure 1 An example of the sample collection container of the present invention is shown. The sample collection container 1 includes a cap-shaped sample collection container upper part 2 and a sample collection container lower part 3 with a capacity of 50 ml, both of which are airtightly joined by screwing. Two connecting parts, ie, a scope-side connecting part 4 and an aspirator-side connecting part 5, are provided on the upper part 2 of the sample collection container, and can be airtightly connected to a suction duct of an endoscope apparatus. In addition, 0.5 ml of 0.5 M EDTA (Wako Pure Chemical Industries, Ltd. K.K. special grade product) was sealed in the sample collection container 1 .

Embodiment 2

[0090] Example 2: Preparation of Specimen

[0091] Remove the suction tube connected to the connection part from the suction bottle of the endoscope device (EVIS LUCERA, Olympus Corporation), connect it to the scope-side connection part of the above-mentioned sample collection container, and simultaneously collect the sample The suction side connection part of the container is connected with the scope side connection part of the suction bottle to form a closed circuit.

[0092] Dilute 4 ml of simethicone (Gascon (registered trademark) drops, Kissei Pharmaceutical Co., Ltd.) (containing 80 mg of dimethyl polysiloxane) with 50 to 100 ml of ordinary water, and dissolve 20,000 units in the diluted solution Pronase (registered trademark) (Pronase (registered trademark) MS, Kakuken Pharmaceutical Co., Ltd.) and 1 g of baking soda were administered to human subjects suffering from various stomach problems, and after 10 to 15 minutes, the subjects were placed in a supine position. ...

Embodiment 3

[0095] Example 3: Evaluation of DNA Quality and Quantity

[0096] i) Evaluation of DNA quality

[0097] The quality of DNA contained in the above samples derived from human subjects with gastric cancer was evaluated by electrophoresis. In the electrophoresis method, 1-2 μg of DNA sample was dissolved in 10 μl of H 2 Diluted in O, run electrophoresis on 1% agarose gel at 100V for 30 minutes. The results of electrophoresis evaluation were as follows: figure 2 shown. In the figure, the left lane is a positive control (1Kb DNA Extension Ladder, Cat. No. 10511-012, Invitrogen), the middle lane is a serum sample, and the right lane is a gastric mucosa washing solution. Either result showed that gastric mucosal wash samples contained DNA of better quality than serum samples and comparable to biopsy samples.

[0098] ii) Evaluation of DNA amount

[0099] The recovered amount of DNA contained in samples derived from human subjects with gastric cancer was evaluated spectrophoto...

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Abstract

The invention relates to a sample containing a gastric mucosa lavage fluid collected from a subject who has received a gastric mucus removal treatment and a method for detecting a disease-related marker using the same. By using the sample of the invention, the disease-related marker can be detected conveniently, less invasively, highly sensitively and highly accurately.

Description

technical field [0001] The present invention relates to a method for detecting disease-related markers, and more specifically to the above-mentioned method using gastric mucosal washings, a specimen containing gastric mucosal washings, a pretreatment agent used in the method, and an instrument used in the method . Background technique [0002] Cancer has been the number one cause of death in Japan for nearly two decades. When looking at cancers that are the cause of death by site, gastric cancer has always been at the top, and the number of gastric cancer cases is variable. In the past, stage III and IV, which represented advanced gastric cancer, occupied the main patient stratum, but now more than 50% are occupied by stage I . Stage I includes Stage Ia and Ib, but all of them have the characteristic that the cancer resides in the submucosa, and are classified as so-called early cancers. [0003] As the treatment guideline for the above-mentioned cancers, less invasive tr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12M1/00C12M1/26C12M3/00G01N33/15C12N15/09
CPCC12Q1/6806C12N15/1003G01N33/15
Inventor 渡边嘉行丰田实今井浩三筱村恭久伊东文生时野隆至
Owner SAPPORO MEDICAL UNIVERSITY
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