Latex immunoturbidimetry pepsinogen I detection kit for eliminating chyle interference

A pepsinogen and latex immunization technology is applied in the field of serum pepsinogen I detection kits, which can solve the problems of increasing the workload of operators, increasing operating costs, and difficulty in arranging clinical testing organizations, and achieves strong chyle interference capability and saves money. Human, clinically significant results

Active Publication Date: 2014-04-02
北京万泰德瑞诊断技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in large-scale population census, health check-up screening, and clinical patient testing, a large number of turbid samples such as chyle and hemolysis are often encountered. At this time, by purchasing sample processing ...

Method used

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  • Latex immunoturbidimetry pepsinogen I detection kit for eliminating chyle interference
  • Latex immunoturbidimetry pepsinogen I detection kit for eliminating chyle interference
  • Latex immunoturbidimetry pepsinogen I detection kit for eliminating chyle interference

Examples

Experimental program
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Effect test

Embodiment 1

[0032] 1. Preparation of pepsinogen I detection kit for eliminating chyle interference

[0033] Preparation of reagent 1: in 50mmol / L, pH7.5 glycine-NaOH buffer, add 20KU / L lipase, 1.2% sodium chloride, 0.3% Tween 20, 5% PEG-6000, 0.5% BSA , 0.05% Proclin300, stir evenly and adjust the pH to 8.0 to obtain the pepsinogen I detection kit reagent 1 which eliminates the interference of chyle.

[0034] Preparation of Reagent 2:

[0035] (1) Antibody cross-linking: Dilute 0.5 mg of mixed duck anti-human pepsinogen Ⅰ IgY antibody with a ratio of 5:1 with 5 ml of 20 mmol / L HEPES-NaOH buffer, then add surface modified carboxyl groups with a diameter of 150 nm. 1.5 mg of EDAC was added, and the reaction was carried out at 37 °C for 8 h, and 0.5 ml of 100 mmol / L Tris-HCl buffer was added, and the reaction was terminated after stirring for 1 h;

[0036] (2) Latex washing: Centrifuge to remove supernatant to remove excess antibody, cross-linking agent and by-products of cross-linking rea...

Embodiment 2

[0087] 1. Preparation of pepsinogen Ⅰ detection kit to eliminate chyle interference

[0088] Preparation of Reagent 1: Add 5KU / L lipase, 4.5% potassium chloride, 0.6% polyoxyethylene phenyl ether, 2% PEG-6000, 2.5% EDTA, 0.08% sodium azide, stirred evenly and adjusted the pH to 7.2, and the pepsinogen Ⅰ detection kit reagent 1 for eliminating chyle interference was obtained.

[0089] Preparation of Reagent 2:

[0090] (1) Antibody crosslinking: After diluting 0.5mg of monoclonal antibody / polyantibody ratio of 6.5:1 mixed rabbit anti-human pepsinogen Ⅰ IgG antibody with 5ml of 10mmol / L MOPSO-HCl buffer, add surface-modified amino groups with a diameter of 200nm Add 2.0mg of NHS to the polystyrene latex solution, react at 37°C for 8h, add 0.5ml of 80mmol / L glycine-NaOH buffer and stir for 1h to terminate the reaction;

[0091] (2) Latex cleaning: Centrifuge to remove the supernatant to remove excess antibodies, cross-linking agents and by-products of the cross-linking reaction...

Embodiment 3

[0129] 1. Preparation of pepsinogen Ⅰ detection kit to eliminate chyle interference

[0130] Preparation of reagent 1: Add 4% polyethylene glycol-magnesium sulfate inulin, 2.5% magnesium sulfate, 1% Tween 80, 6% PEG- 8000, 5% sorbitol, and 0.1% Proclin300, the pepsinogen I detection kit reagent 1 that eliminates chyle interference was obtained.

[0131] Preparation of Reagent 2:

[0132](1) Antibody cross-linking: After diluting 0.5 mg of monoclonal antibody / polyantibody ratio of 9:1 mixed chicken anti-human pepsinogen II IgY antibody with 5 ml of 50 mmol / L phosphate buffer, add surface-modified chloromethyl, diameter 250nm polystyrene latex solution, then add 1.5mg EDAC, react at 37°C for 8h, add 0.5ml80mmol / L Tris-HCl buffer and stir for 1h, then terminate the reaction;

[0133] (2) Latex cleaning: Centrifuge to remove the supernatant to remove excess antibodies, cross-linking agents and by-products of the cross-linking reaction, etc., and the bottom precipitate is the coa...

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Abstract

The invention relates to the technical field of medical examination, and particularly relates to a latex immunoturbidimetry serum pepsinogen I detection kit for eliminating chyle interference. The kit provided by the invention comprises (1) a pepsinogen I calibrator; (2) a reagent 1 with a preset chyle remover; and (3) a reagent 2 containing the monoclonal antibody and polyclonal antibody coated latex particles against human pepsinogen I. According to the kit provided by the invention, the combination reaction between a substrate to be detected in a sample and a specific antibody in the reagent is amplified through the latex agglutination effect; with the given wavelength, the turbidity formed by the reaction is related to the content of the substrate to be detected, thus the content of the substrate to be detected is calculated. The kit provided by the invention is used for detecting the content of pepsinogen I in human serum, has high sensitivity and good specificity, and can eliminate chyle interference in the sample; moreover, the kit is simple and quick to operate and has high practicability and wide application range.

Description

technical field [0001] The invention belongs to the technical field of medical examination, and relates to a serum pepsinogen I detection kit for latex immunoturbidimetric method eliminating chyle interference. Background technique [0002] Pepsinogen (PG) is an inactive precursor of pepsin in gastric juice, a precursor of aspartic protease secreted by gastric mucosa, and a single-chain polypeptide with a molecular weight of 42,000 Da, which is transformed into active protease in the stomach. Pepsin. [0003] According to biochemical properties, immunogenicity, cell origin and distribution in tissues, pepsinogen can be divided into two subgroups: pepsinogen Ⅰ (PGⅠ) and pepsinogen Ⅱ (PGⅡ). , called PG I (PGA), mainly secreted by the principal cells of the fundic gland and cervical mucus cells; 6-7 components are called PG II (PGC), in addition to being secreted by the above two cells, the mucus cells of the pyloric gland, Cardiac glands and Brunner glands in the upper part ...

Claims

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Application Information

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IPC IPC(8): G01N33/577
CPCG01N33/68G01N2333/96477
Inventor 李雪刘红春欧阳卓君
Owner 北京万泰德瑞诊断技术有限公司
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