Chemiluminescent quantitative determination kit for pepsinogen II and preparation method of chemiluminescent quantitative determination kit

A quantitative detection technology for pepsinogen, applied in pepsinogen Ⅱ chemiluminescent quantitative detection kit and its preparation field, can solve the problem of no correlation of gastric acid secretion and achieve high sensitivity

Inactive Publication Date: 2015-04-29
HENAN MAINCARE BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In H. pylori-negative subjects, the level of PG I was significantly correlated with the amount of gastric

Method used

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  • Chemiluminescent quantitative determination kit for pepsinogen II and preparation method of chemiluminescent quantitative determination kit
  • Chemiluminescent quantitative determination kit for pepsinogen II and preparation method of chemiluminescent quantitative determination kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 Preparation of Pepsinogen II Chemiluminescent Quantitative Detection Kit by Using Carbonate Coating Buffer

[0051] 1. Preparation of PBST solid lotion and diluent:

[0052] 1) The formula of PBST solid lotion is

[0053] K H 2 PO 4 4.8g

[0054] Na 2 HPO 4 •12H 2 O 28.8g

[0055] NaCl 155g

[0056] Tween-20 10mL.

[0057] 2) The mixed PBST solid lotion is dried under reduced pressure and low temperature under the condition of air pressure 500Pa, temperature 10°C and humidity 30%. After 1 hour, it is packed in aluminum foil bags. When using, every 5g of PBST solid lotion is dissolved in 500mL deionized Mix in water to prepare PBST solid lotion dilution.

[0058] 2. Preparation of coated microplates:

[0059] 1) Selection of antibody: use mouse monoclonal antibody.

[0060] 2) Coating of microwell plates: Dilute the pepsinogen II monoclonal antibody to a concentration of 3 μg / mL with 0.02 mol / L carbonate buffer solution with a pH value of 9.6 t...

Embodiment 2

[0100] Example 2 Preparation of Pepsinogen II Chemiluminescence Quantitative Detection Kit by Using Phosphate Coating Buffer

[0101] Coating: Dilute the pepsinogen II monoclonal antibody to a concentration of 2 μg / mL with 0.2 mol / L phosphate coating buffer with a pH value of 6.5, prepare a coating solution, and absorb it at 100 μL / well Coated on a microwell plate at 37°C for 4h. Specifically, the preparation method of the phosphate coating buffer with a pH value of 6.5 is: 68.5 mL of 0.2 mol / L NaH 2 PO 4 solution and 31.5mL of 0.2mol / L Na 2 HPO 4 The solution is mixed.

[0102] Rabbit-derived human pepsinogen II monoclonal antibody was selected as the antibody, and the preparation method of other reagents and components was the same as that in Example 1.

Embodiment 3

[0103] Example 3 Preparation of pepsinogen Ⅱ chemiluminescence quantitative detection kit of the present invention by using Tris-HCl coating buffer

[0104] Coating: Dilute the pepsinogen II monoclonal antibody to a concentration of 0.5 μg / mL with 0.05 mol / L Tris-HCl coating buffer with a pH value of 7.4 to prepare a coating solution, and add 100 μL / well It was adsorbed on a microwell plate and coated at 37° C. for 4 hours. Specifically, the preparation method of the Tris-HCl coating buffer with a pH value of 7.4 is as follows: after mixing 50.0 mL of 0.1 mol / L Tris alkali solution and 42.0 mL of 0.1 mol / L HCl solution, add water to volume to 100mL.

[0105] The preparation method of other reagents and components is consistent with the method in Example 1.

[0106] The using method of kit of the present invention

[0107] 1. Preparation before the experiment:

[0108] 1) Take the kit out of the refrigerated environment and place it at room temperature (18°C~25°C) to ...

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Abstract

The invention relates to the field of immunodetection, in particular to a chemiluminescent quantitative determination kit for pepsinogen II and a preparation method of the chemiluminescent quantitative determination kit. The chemiluminescent quantitative determination kit for the pepsinogen II comprises (1) a pepsinogen II antibody coated micropore plate, (2) an HRP-labeled pepsinogen II antibody, (3) series pepsinogen II calibration materials diluted by a calibration material diluent, (4) a luminescent substrate A, (5) a luminescent substrate B and (6) a solid washing liquid. The kit disclosed by the invention is capable of detecting the content of the pepsinogen II in serum; and the level of the pepsinogen II in the serum is in positively relevant to atrophic gastritis and peptic ulcer; in treatment of peptic ulcer, the treatment result can be judged by monitoring the change of the degermed pepsinogen; and the chemiluminescent quantitative determination kit is noninvasive, simple and fast in detection method, high in sensitivity, wide in detection range and the like.

Description

technical field [0001] The invention relates to the field of immune detection, in particular to a pepsinogen II chemiluminescence quantitative detection kit and a preparation method thereof. Background technique [0002] Pepsinogen (PG) is a digestive enzyme precursor secreted by the stomach and an inactive precursor of pepsin in gastric juice. It is a protein polypeptide chain composed of 375 amino acids with an average molecular weight of 42,000. There are seven groups of pepsin isoenzymes in the gastric mucosa. PG is synthesized on ribosomes, secreted out of cells by Golgi apparatus, and becomes pepsin after being activated by hydrochloric acid. According to biochemical properties, immunogenicity, cell source and tissue distribution, it can be divided into two subgroups, PGⅠ and PGⅡ, of which 6~7 components have similar immunogenicity and are called PGⅡ. In addition to the secretion of the principal cells of the gland, the mucous neck cells of the oxyntic glands, the mu...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/535G01N21/76
CPCG01N33/577G01N21/76G01N33/535
Inventor 赵焕朝俞宁胡锰鸽翟秋菊张喜娟
Owner HENAN MAINCARE BIOLOGICAL TECH
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