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Pepsinogen II (PGII) quantitative assaying kit, preparing method thereof and detecting method thereof

A technology for quantitative determination of pepsinogen, applied in the field of biological immunodiagnosis of medical devices, can solve the problems of unfavorable high-throughput automatic detection, poor specificity, low sensitivity, etc., and achieves optimized chemiluminescence enhancement system, small variation , the effect of high sensitivity

Inactive Publication Date: 2016-03-02
JIANGSU ZECEN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This technique has low sensitivity, narrow linearity, poor specificity, and is not conducive to high-throughput automatic detection

Method used

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  • Pepsinogen II (PGII) quantitative assaying kit, preparing method thereof and detecting method thereof
  • Pepsinogen II (PGII) quantitative assaying kit, preparing method thereof and detecting method thereof
  • Pepsinogen II (PGII) quantitative assaying kit, preparing method thereof and detecting method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] The configuration of various buffers is as follows:

[0056] 1. Tris salt pH8.0 buffer

[0057] Tris: 12.12g, sodium chloride 5.82g, add to 1L of purified water, stir well until completely dissolved, adjust the final pH to 7.5 with hydrochloric acid.

[0058] 2. Preparation of calibrator buffer

[0059] Add 0.01g of tetracycline and 0.1g of neomycin sulfate to 1L of neonatal bovine serum, fully dissolve and process through a 0.22 μm filter membrane to obtain.

[0060] 3. Anti-reagent buffer

[0061] Tris: 12.12mg~60.57mg, tetracycline: 0.01g~0.05g, sheep serum: 1g~5g, newborn bovine serum 3g~10g, horse serum 1g~5g, add 1L purified water, stir well until completely dissolved;

[0062] 4. Magnetic particle buffer

[0063] Tris: 12.12mg, sodium chloride 5.82mg, methyl cellulose ether 50g, add to 1L of purified water, stir well until completely dissolved.

[0064] 5. Luminescence substrate buffer

[0065] Tris12.12g~121.14g, sodium chloride 5.82g, lucigenin 0.03g, add...

Embodiment 2

[0068] Embodiment 2: the preparation of the quantitative assay kit of pepsinogen II (PGII)

[0069] 1. Preparation of calibrators and quality controls

[0070] First: Dissolve pepsinogen II (PGII) with a standard buffer solution, and configure it as a calibrator and a quality control product with a target concentration as shown in Table 1; the pepsinogen II (PGII) used in this example is purchased from home and abroad Well-known manufacturers fitzgerald company.

[0071] Table 1: Preparation of calibrators and controls

[0072]

[0073] 2. The preparation method of the anti-reagent is as follows:

[0074] (1), fluorescein isothiocyanate is coupled with pepsinogen II (PGII) antibody to obtain fluorescein isothiocyanate-labeled pepsinogen II (PGII) coated antibody:

[0075] First, use anti-reagent buffer to prepare fluorescein isothiocyanate solution with a concentration of 2.5mg / mL fluorescein isothiocyanate, according to the mass ratio of pepsinogen II (PGII) to fluoresc...

Embodiment 3

[0087] Embodiment 3: the step that detects pepsinogen II (PGII) with pepsinogen II (PGII) quantitative assay kit

[0088] Utilize this pepsinogen II (PGII) quantitative assay kit to detect the method for pepsinogen II (PGII), the method comprises steps:

[0089] (1) Take three test tubes and add 30 μL pepsinogen II (PGII) calibrator, 30 μL pepsinogen II (PGII) quality control, and 30 μL test sample;

[0090] (2) Add 60 μL of anti-reagent to each test tube, cover the test tube with a plastic film, shake the test tube gently for 30 s, and place it in a water bath at 37°C for 15 minutes;

[0091] (3) Add 30 μL of magnetic particle reagent to each test tube, cover the test tube with plastic film, shake the test tube gently for 30 seconds, and place it in a water bath at 37°C for 5 minutes;

[0092](4) Precipitate the test tube on the magnetic separator for 2 minutes, slowly invert the test tube and the magnetic separator, pour out the supernatant, put the inverted test tube toget...

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Abstract

The invention discloses a pepsinogen II (PGII) quantitative assaying kit. The kit comprises a calibrated material, a quality control material, a resistance reagent, a magnetic particle reagent and a light emitting substrate. The invention further discloses a preparing method of the kit and a method for detecting PGII through the kit. According to the kit, the preparing method and the method, the resistance reagent is prepared from a PGII coated antibody labeled through fluorescein isothiocyanate and a PGII labeled antibody labeled through alkaline phosphatase, the magnetic particle reagent is prepared in the mode that the fluorescein isothiocyanate resistant antibody is coupled with carboxyl magnetic beads, even mixing and separation of an immunoreaction are easier accordingly, and the reaction speed is greatly increased; the novel chemical light emitting substrate APLS serves as the substrate, and therefore the sensitivity and the specificity of the kit are improved. The detecting kit is reliable in performance, high in sensitivity, wide in linear range and capable of being used in cooperation with semi-automatic and full-automatic instruments.

Description

technical field [0001] The invention belongs to the field of biological immune in vitro diagnosis of medical devices, and mainly relates to a kit for quantitatively detecting pepsinogen II (PGII) in blood based on magnetic particle separation chemiluminescence and a preparation method thereof, and the quantitative detection of blood using the kit The method of pepsinogen II (PGII). Background technique [0002] Pepsinogen is the precursor of pepsin. According to its biochemical properties and immunogenicity, it is divided into 2 subgroups. The immunogenicity of components 1-5 is the same, called pepsinogen I, which is mainly composed of fundic glands. Secreted by chief cells and mucus neck cells; components 6 and 7 are called pepsinogen II, in addition to being secreted by chief cells and mucus neck cells of fundic glands, mucus neck cells of cardia glands and pyloric glands of gastric antrum, and ten The upper duodenum also produces pepsinogen II. [0003] Under normal ci...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/531G01N33/533G01N21/76
Inventor 夏振伟杨旻汪丹
Owner JIANGSU ZECEN BIOTECH CO LTD
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