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Reagent kit and method for measuring concentration of glycocholic acid

A technology of glycocholic acid and a kit, which is applied in the field of kits for determining the concentration of glycocholic acid, can solve the problems of poor anti-interference performance, high price, and cannot be used for automatic analysis, and achieves improved stability, improved accuracy, and eliminated the Effects of non-specific cross-reactivity

Active Publication Date: 2016-10-05
上海微鸿企业管理有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The technical problem to be solved by the present invention is that in order to overcome the existing high-pressure liquid chromatography and enzyme-linked immunoassay that require special measuring instruments, the operation is complicated, the test takes a long time, it cannot be applied to the automatic analysis of clinical large flow, and the price is too high. Expensive; and if the immune turbidimetric assay method is used mechanically, the detection accuracy of glycocholic acid is low, and the defect of anti-interference performance is poor, a kind of test kit and method for measuring glycocholic acid concentration are provided

Method used

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  • Reagent kit and method for measuring concentration of glycocholic acid
  • Reagent kit and method for measuring concentration of glycocholic acid

Examples

Experimental program
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Effect test

Embodiment 1

[0042] (1) Preparation of Glycocholic Acid-Ovalbumin Conjugate

[0043] Add 10 mg of ovalbumin to 1.0 mL of 25 mmol / L MES (2-morpholineethanesulfonic acid) buffer solution with a pH of 6.1, stir rapidly, add glycocholic acid, and slowly drop in 60 mmol / L EDAC (1-(3- 300 μL of dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride) solution was stirred rapidly for 2 hours, and 2.5 mL of 50 mmol / L Tris buffer solution with a pH of 8.2 was added to terminate the reaction for 30 minutes. Dialyze at 4°C against citrate buffer with pH 6.1 for later use.

[0044] (2) Preparation of Reagent 1

[0045] Take 2 mL of latex particles with a concentration of 10%, carboxyl groups on the surface, and a diameter of 100 nm (CB0100E product of Polymicrospheres Company), and add it to 5 mL of 20 mM MES buffer solution with pH 6.1. While stirring, add 300 mg of Sulfo-NHS and 30 mg of EDAC , react at room temperature for 20 minutes, centrifuge to remove the supernatant, then add 35 mL of pH 8.35...

Embodiment 2

[0063] (1) Preparation of glycocholic acid-hemocyanin conjugate

[0064] Add 10 mg of hemocyanin to 1.0 mL of 20 mmol / L MES (2-morpholineethanesulfonic acid) buffer solution with a pH of 6.1, stir rapidly, add glycocholic acid, and slowly drop in 50 mmol / L EDAC (1-(3- 300 μL of dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride) solution was stirred rapidly for 2 hours, and 2.5 mL of 50 mmol / L Tris buffer solution with a pH of 8.2 was added to terminate the reaction for 30 minutes. Dialyze with MES buffer at pH 6.1 at 4°C for later use.

[0065] (2) Preparation of Reagent 1

[0066]Take 2mL of CB0213E latex particles from Polymicrospheres company with a concentration of 10% and carboxyl groups on the surface, and add them to 5mL of 20mM MES buffer solution with pH 6.1. While stirring, add 300mg of Sulfo-NHS and 30mg of EDAC, react at room temperature for 20 minutes, and centrifuge to remove The supernatant was then added to 35 mL of pH 8.350 mM boric acid buffer solution...

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Abstract

The invention discloses a reagent kit and method for measuring the concentration of glycocholic acid. The reagent kit comprises a reagent (1) prepared from latex particles coated with a glycocholic acid resisting monoclonal antibody, and a reagent (2) prepared from latex particles coated with glycocholic acid-inert carrier conjugates. When used for detecting glycocholic acid, the reagent kit can easily enhance immunity turbidity and amplify detection signals, has the advantages of being free of special instruments, easy and convenient to operate, high in sensitivity and low in cost, is capable of quickly measuring large-flow samples clinically and has extremely high economic value, and it becomes possible that glycocholic acid becomes a clinical conventional detection item.

Description

technical field [0001] The invention belongs to the technical field of substance inspection and determination, and in particular relates to a kit and a method for determining the concentration of glycocholic acid. Background technique [0002] Cholyglycine (CG) is one of the conjugated cholic acids formed by the combination of bile acid and glycine. The normal metabolic pathway of CG is the entero-hepatic circulation, which is synthesized by liver cells, discharged into the gallbladder through the capillary and bile ducts, and enters the duodenum with bile to help digest food. 95% of bile acid is reabsorbed in the terminal ileum, and then returned to the liver through the portal vein, where it is taken up and reused by liver cells. In the serum, it mainly exists in the form of protein binding, and the total amount spilled into the systemic circulation is less than 1%. Under normal circumstances, the bile acid content in peripheral blood is very small, and the serum CG conc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
Inventor 不公告发明人
Owner 上海微鸿企业管理有限公司
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