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Method for detecting expression quantity of carcinoembryonic antigen mRNA and special kit thereof

A carcinoembryonic antigen and kit technology, applied in biochemical equipment and methods, microbiological determination/inspection, etc., can solve the problems of low positive rate, lack of detection technology, and incapable of quantitative analysis, etc., to achieve accuracy and specificity Good, high-sensitivity effect

Active Publication Date: 2010-03-17
UNIV OF SCI & TECH OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current detection methods have the problem of low positive rate, and there is no suitable detection technology. The ordinary RT-PCR detection can only detect the expression of CEA mRNA in the peripheral blood of patients with gastrointestinal tumors qualitatively, and cannot perform quantitative analysis.

Method used

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  • Method for detecting expression quantity of carcinoembryonic antigen mRNA and special kit thereof
  • Method for detecting expression quantity of carcinoembryonic antigen mRNA and special kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1, composition and preparation of kit

[0047] The composition of the kit and its preparation method are as follows:

[0048] 1. Cell lysate I

[0049] It consists of tris(hydroxymethyl)aminomethane hydrochloride with a final concentration of 0.1mol / L, sodium chloride with a final concentration of 0.1mol / L and magnesium chloride with a final concentration of 0.05mol / L; the solvent is water. Wherein, the final concentration is the concentration of each substance in cell lysate I.

[0050] Cell lysate I has a pH value of 7.6 at 25°C.

[0051] 2. Cell Lysis Solution II

[0052] It consists of guanidine isothiocyanate with a final concentration of 3mol / L, guanidine hydrochloride with a final concentration of 2mol / L, sodium acetate with a final concentration of 0.3mol / L and sodium lauryl sulfate with a final concentration of 0.2%. It is water. Wherein, the final concentration is the concentration of each substance in the cell lysate II.

[0053] The cell lys...

Embodiment 2

[0182] Embodiment 2, detection of carcinoembryonic antigen gene mRNA expression

[0183] The kit prepared in Example 1 was used to detect the expression of carcinoembryonic antigen gene mRNA in the specimens of the following experimental group and control group.

[0184] Experimental group: 15 patients with pathologically diagnosed gastric cancer, 5 of whom were clinically diagnosed with metastasis; 10 patients with pathologically diagnosed esophageal cancer, 4 of whom were clinically diagnosed with metastasis; 8 patients with pathologically diagnosed colon cancer, Three of them were clinically confirmed to have metastases.

[0185] Control group: 10 patients with colon polyps, 10 patients with gastric ulcer and 10 healthy people.

[0186] All samples were obtained with the consent of the subjects.

[0187] 1. Experiment preparation

[0188] 1. Dilute the cell lysate I with sterilized deionized water at a ratio of 1:9 to obtain the cell lysate I dilution.

[0189] 2. Add 2...

Embodiment 3

[0239] Embodiment 3, test kit and application thereof

[0240] 1. Kit composition and preparation method

[0241] Same as described in Example 1, the difference is as follows:

[0242] 1. PCR reaction buffer: composed of tris(hydroxymethyl)aminomethane hydrochloride with a final concentration of 18.0mmol / L, magnesium chloride with a final concentration of 2.60mmol / L and potassium chloride with a final concentration of 90.5mmol / L , the solvent is water; wherein, the final concentration is the final concentration of each substance in the PCR reaction buffer.

[0243] 2. PCR reaction solution: consisting of the PCR reaction buffer, Taq enzyme, dATP, dCTP, dGTP, dTTP, dUTP, the primer pair shown in sequence 1 and 2 in the sequence listing, the probe shown in sequence 3 in the sequence listing and urine Pyrimidine DNA glycosylase, the rest is water;

[0244] The concentration of the PCR reaction buffer in the PCR reaction solution is 0.9 μl / μl;

[0245] The concentration of the...

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Abstract

The invention discloses a method for detecting the expression quantity of carcinoembryonic antigen mRNA and a special kit thereof. The kit comprises a primer pair: the sequence of one primer is shownby sequence 1 in a sequence table, and the sequence of the other primer is shown by sequence 2 in the sequence table. The kit and the method provided by the invention can be used for accurately quantifying the expression quantity of the carcinoembryonic antigen mRNA in tissue to be detected; and for instance, the invention can be used for carrying out qualitative and quantitative analysis on the expression situation of CEA mRNA in peripheral blood and bone marrow of tumor patients, and has important significance for judging the occurrence, the transfer, the recrudescence, the treatment effectevaluation and the dynamic observation of illness state of gastric cancer, esophagus cancer, colorectal cancer and rectal cancer. Experiment shows that the specificity of the detection method can reach 100% and the high sensitivity thereof can reach 100 copies / ml whole blood. Therefore, the invention plays an important role in the field of medical test.

Description

technical field [0001] The invention relates to a method and a special kit for detecting the expression level of carcinoembryonic antigen mRNA. Background technique [0002] Gastrointestinal tumors, including esophageal cancer, gastric cancer, rectal cancer, colon cancer, etc., are common malignant tumors that endanger human health. Due to differences in geographical environment and living habits, the incidence of various tumors varies greatly across the world. Taking esophageal cancer as an example, about 300,000 people worldwide die from esophageal cancer every year. my country is an area with a high incidence of esophageal cancer. About 150,000 people die from esophageal cancer every year, accounting for nearly 1 / 4 of the total number of deaths from malignant tumors. The incidence of esophageal cancer in my country has obvious regional distribution characteristics. For example, the Taihang Mountains in North China and the northwestern Sichuan Basin are distributed in irr...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 程民魏海明田志刚
Owner UNIV OF SCI & TECH OF CHINA
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