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Kit for detecting gastrin-17 and preparation method as well as application thereof

A gastrin and kit technology, applied in biological testing, measuring devices, material inspection products, etc., can solve problems such as inaccurate test results, lower reagent specificity and sensitivity, complex gastrin components, etc.

Active Publication Date: 2015-05-13
SHENZHEN NEW INDS BIOMEDICAL ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although detecting the full fragment of gastrin can prevent missed detection when detecting the clinical gastrin content, the components of gastrin are too complicated, and not every component can be used for the detection and diagnosis of clinical diseases; and in the detection of gastrin The polyantibody used for the whole fragment will seriously reduce the specificity and sensitivity of the reagent, resulting in inaccurate detection results

Method used

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  • Kit for detecting gastrin-17 and preparation method as well as application thereof
  • Kit for detecting gastrin-17 and preparation method as well as application thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0063] The following is the preparation method of each component of the kit:

[0064] Preparation 1: Preparation of magnetic sphere suspension coated with the first anti-gastrin-17 antibody

[0065] The immunomagnetic spheres used in this preparation step are selected from the nano-magnetic microsphere suspension produced by Merck Company with a concentration of 100 mg / ml and a hydroxyl group of 95 mg KOH / g.

[0066] (1) Preparation of buffer:

[0067] Weigh 2.55g of sodium acetate trihydrate, dissolve it in 4500ml of purified water, add 14ml of acetic acid, and mix well to obtain an acetate buffer solution with a pH of 3.6.

[0068] (2) Magnetic microsphere connection (magnetic microsphere connection CMC method):

[0069] Suspend the magnetic microspheres in the above-mentioned pH3.6 acetate buffer solution 5 times of the coating volume, so that the concentration of the magnetic beads is 20mg / ml; then add 1-cyclohexyl-2-morpholinoethylcarbodiimide to Tosylate (CMC) to a co...

Embodiment 1

[0145] Preparation of kit components:

[0146] Prepare the ABEI solution labeled with the first anti-gastrin-17 antibody according to the above preparation 4;

[0147] According to the above-mentioned preparation 1, the magnetic sphere suspension coated with the second anti-gastrin-17 antibody was prepared;

[0148] Prepare gastrin-17 low point calibrator and high point calibrator according to Preparation 12 above.

[0149] Using gastrin-17 standard substance, using bovine serum as a solvent, prepare 10 parts of standard solution with different concentrations, and use the kit components prepared in this example to detect the 10 parts of the solution. The detection steps are as follows:

[0150] 1) Add 40 μl of the solution to be tested, the high-point calibrator and the low-point calibrator into different cuvette wells;

[0151] 2) Add 100 μl of ABEI solution labeled with the first anti-gastrin-17 antibody;

[0152] 3) Add 20 μl of magnetic ball suspension coated with the s...

Embodiment 2

[0159] Preparation of kit components:

[0160] Prepare the ABEI solution labeled with the first anti-gastrin-17 antibody according to the above preparation 4;

[0161] According to the above preparation 3, a magnetic sphere suspension coated with goat anti-FITC polyclonal antibody was prepared;

[0162] Prepare the FITC solution labeled with the second anti-gastrin-17 antibody according to the above-mentioned preparation 6;

[0163] Prepare gastrin-17 low point calibrator and high point calibrator according to Preparation 12 above.

[0164] Using the detection kit components of this embodiment to carry out the method for gastrin-17 chemiluminescent immunoassay, with reference to the detection method in embodiment 1, 10 parts of standard solution in embodiment 1 and 5 cases of confirmed duodenal Serum samples from patients with intestinal ulcer were tested, and the test results are shown in Table 1 and Table 2, respectively.

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Abstract

The invention provides a kit for detecting gastrin-17 and a preparation method as well as application thereof. The kit comprises components A and B, wherein the component A is a first anti-gastrin-17 antibody marked with a tracing marker or coated with a magnetic ball, and the component B is a second anti-gastrin-17 antibody coated with the magnetic ball or marked with the tracing marker, or the gastrin-17; moreover, any one of the components A and B is marked with the tracing marker, the other one is coated with the magnetic ball; and the first anti-gastrin-17 antibody and the second anti-gastrin-17 antibody have different binding sites with the gastrin-17. The invention provides a method for detecting the gastrin-17, with the kit provided by the invention, content of the gastrin-17 in a sample can be detected accurately and sensitively according to a double antibody sandwich method principle or a competition method principle.

Description

technical field [0001] The invention relates to the technical field of biological substance detection, in particular to a kit for detecting gastrin-17, a preparation method thereof, and a method for detecting gastrin-17 using the kit. Background technique [0002] Gastrin is an important gastrointestinal hormone, which is secreted by G cells (gastrin containing cells) present in the pyloric mucosa of the stomach, and then enters the blood and then acts on the stomach to promote the secretion of gastric juice. G cells are mainly distributed in the antrum of the stomach, and during renal metabolism, serum gastrin increases when renal insufficiency occurs. The pyloric antrum of the stomach is mechanically stimulated by food, etc., and gastrin is secreted when the pH of the pylorus is alkaline. As a result, the secretion of gastric juice, especially hydrochloric acid, becomes vigorous. When the pH inside the stomach decreases, the secretion of hydrochloric acid and gastrin sto...

Claims

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Application Information

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IPC IPC(8): G01N33/74G01N33/532G01N21/76
CPCG01N33/74G01N2333/595
Inventor 饶微孙普李昀轩刘望杨雅丽袁锦云李婷华
Owner SHENZHEN NEW INDS BIOMEDICAL ENG
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