Latex immunoturbidimetry type pepsinogen II detection kit capable of eliminating chyle interference

A pepsinogen and latex immunization technology, which is applied in the field of serum pepsinogen II detection kits, can solve the problems of clinical inspection organization and arrangement, increase the workload of operators, increase operating costs, etc., and achieve strong chyle interference ability and clinical Significant and labor-saving effect

Active Publication Date: 2014-04-09
北京万泰德瑞诊断技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 type pepsinogen II detection kit capable of eliminating chyle interference
  • Latex immunoturbidimetry type pepsinogen II detection kit capable of eliminating chyle interference
  • Latex immunoturbidimetry type pepsinogen II detection kit capable of eliminating chyle interference

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] 1. Preparation of pepsinogen Ⅱ detection kit to eliminate chyle interference

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

[0034] Preparation of Reagent 2:

[0035] (1) Antibody cross-linking: After diluting 0.5 mg of the mixed duck anti-human pepsinogen II IgY antibody with a monoclonal antibody / polyantibody ratio of 5:1 with 5 ml of 20 mmol / L HEPES-NaOH buffer, add surface-modified carboxyl groups with a diameter of 150 nm Add 1.5 mg of EDAC to the polystyrene latex solution, react at 37°C for 8 h, add 0.5 ml of 100 mmol / L Tris-HCl buffer and stir for 1 h to terminate the reaction;

[0036] (2) Latex cleaning: Centrifuge to remove the supernatant to remove excess antibodies, cross-linking agents and by-products of c...

Embodiment 2

[0086] 1. Preparation of pepsinogen Ⅱ detection kit to eliminate chyle interference

[0087] 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 reagent 1 of the pepsinogen II detection kit for eliminating chyle interference was obtained.

[0088] Preparation of Reagent 2:

[0089] (1) Antibody cross-linking: After diluting 0.5 mg of monoclonal antibody / polyantibody ratio of 6.5:1 mixed rabbit anti-human pepsinogen II IgG antibody with 5 ml of 10 mmol / L MOPSO-HCl buffer, add surface-modified amino groups with a diameter of 200 nm 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;

[0090] (2) Latex cleaning: Centrifuge to remove the supernatant to remove excess antibodies, cross-linking agents and by-products of the cross-li...

Embodiment 3

[0128] 1. Preparation of pepsinogen Ⅱ detection kit to eliminate chyle interference

[0129] 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 II detection kit reagent 1 that eliminates chyle interference was obtained.

[0130] Preparation of Reagent 2:

[0131](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;

[0132] (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 co...

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Abstract

The invention relates to the technical field of medical examination and particularly relates to a latex immunoturbidimetry type pepsinogen II detection kit capable of eliminating chyle interference. The kit provided by the invention comprises: (1) a pepsinogen II calibrator; (2) a reagent 1 with a preset chyle remover; and (3) a reagent 2 containing a monoclonal antibody of anti-human pepsinogen II and polyclonal antibody coated latex particles. According to the kit using the method, a conjugation reaction of substance to be detected in a sample and specific antibodies in the reagents is amplified by a latex agglutination effect, turbidity formed by the reaction is associated with the content of the substance to be detected under a given wavelength, so that the content of the substance to be detected can be calculated. The kit provided by the invention is used for detecting the content of pepsinogen II in human serum, and the kit is high in sensitivity, good in specificity, capable of eliminating chyle interference in the sample, quick and convenient to operate, strong in practicability and is wide in application range.

Description

technical field [0001] The invention belongs to the technical field of medical examination, and relates to a serum pepsinogen II detection kit for latex immune turbidimetry to eliminate 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 of th...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/68G01N2333/96477
Inventor 李雪白春洋欧阳卓君
Owner 北京万泰德瑞诊断技术有限公司
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