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Reagent kit for measuring glycocholic acid in human serum and method for applying reagent kit

A technology of glycocholic acid and human serum, applied in the field of immunodetection, can solve the problems of low concentration samples and low sensitivity of small molecule antigens, and achieve the effects of prolonging storage time, shielding interference and simplifying detection steps.

Active Publication Date: 2017-06-13
北京美德美康生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Among them, the existing latex-enhanced immunoturbidimetric method, that is, the direct immunoturbidimetric method, has a simple and fast process, but its sensitivity to low-concentration samples and small molecule antigens is relatively low

Method used

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  • Reagent kit for measuring glycocholic acid in human serum and method for applying reagent kit
  • Reagent kit for measuring glycocholic acid in human serum and method for applying reagent kit
  • Reagent kit for measuring glycocholic acid in human serum and method for applying reagent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Glycocholic acid reagent R1:

[0052]

[0053] After dissolving the above raw materials with 800ml purified water, add purified water to make the volume up to 1000ml, and then adjust the pH to 6.0 with hydrochloric acid and sodium hydroxide.

[0054] Glycocholic acid reagent R2:

[0055] 1. Carboxy latex microspheres: 200nm latex microspheres

[0056] 2. CG antibody concentration: take 20mg / ml as an example for coupling

[0057] S1. Take 5ml of coating buffer (20mmol / L MES, PH6.0), add 80ul of carboxyl latex microspheres, and then add 50ul of CG antibody (rabbit anti-human glycocholic acid antibody), and react at 37°C for 1 hour ;

[0058] S2. Dilute a certain amount of EDC reagent to 1mg / ml with the coating buffer, add 50ul to the reaction solution of S1, and continue the reaction for 1 hour;

[0059] S3. After the reaction is over, use ultrasonic pulverization;

[0060] S4. Centrifuge in a centrifuge, 10,000 rpm, 10 minutes, discard the supernatant, add the same volume of couplin...

Embodiment 2

[0068] Glycocholic acid reagent R1:

[0069]

[0070]

[0071] After dissolving the above raw materials with 800ml purified water, add purified water to make the volume to 1000ml, and then adjust the pH to 6.0 with hydrochloric acid or sodium hydroxide.

[0072] Glycocholic acid reagent R2:

[0073] 1. Carboxy latex microspheres: 300nm latex microspheres

[0074] 2. CG antibody concentration: take 20mg / ml as an example for coupling

[0075] S1. Take 5ml of coating buffer (20mmol / L MES, PH6.0), add 80ul of carboxyl latex microspheres, and then add 50ul of CG antibody (rabbit anti-human glycocholic acid antibody), and react at 37°C for 1 hour ;

[0076] S2. Dilute a certain amount of EDC reagent to 1 mg / ml with the coating buffer, add 50ul to the above reaction solution, and continue the reaction for 1 hour;

[0077] S3. After the reaction is over, use ultrasonic pulverization;

[0078] S4. Centrifuge in a centrifuge, 10,000 rpm, 10 minutes, discard the supernatant, add the same volume of c...

Embodiment 3

[0087] Glycocholic acid reagent R1:

[0088]

[0089] After dissolving the above raw materials with 800ml purified water, add purified water to make the volume to 1000ml, and then adjust the pH to 6.0 with hydrochloric acid or sodium hydroxide.

[0090] Glycocholic acid reagent R2:

[0091] 1. Carboxy latex microspheres: 150nm latex microspheres

[0092] 2. CG antibody concentration: take 20mg / ml as an example for coupling

[0093] S1. Take 5ml of coating buffer (20mmol / L MES, PH6.0), add 80ul of carboxyl latex microspheres, then add 50ul of CG antibody (chicken anti-human glycocholic acid antibody), and react at 37°C for 1 hour ;

[0094] S2. Dilute a certain amount of EDC reagent to 1 mg / ml with the coating buffer, add 50ul to the above reaction solution, and continue the reaction for 1 hour;

[0095] S3. After the reaction is over, use ultrasonic pulverization;

[0096] S4. Centrifuge with a centrifuge at 10,000 rpm for 10 minutes, discard the supernatant, add an equal volume of coupli...

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Abstract

The invention discloses a reagent kit for measuring glycocholic acid in human serum. The reagent kit comprises glycocholic acid reagents R1, glycocholic acid reagents R2 and glycocholic acid standard substances. The reagent kit has the advantages that competition processes are used in detection procedures, accordingly, variation of the turbidity of test solution can be controlled in wide ranges, the glycocholic acid can be detected by the aid of common analytical instruments, and the reagent kit is favorable for guaranteeing the accuracy of the instruments.

Description

Technical field [0001] The invention relates to the field of immunoassays, in particular to a kit for determining glycocholic acid in human serum and an application method thereof. Background technique [0002] Serum glycocholic acid (CG) is one of the secondary bile acids that combine bile acid with glycine. In liver cells, cholesterol is converted into primary bile acid through an extremely complex enzymatic reaction during the period. [0003] The normal metabolic pathway of glycocholic acid (CG) is intestinal-hepatic circulation, and the normal metabolic pathway of CG is intestinal-hepatic circulation. It is synthesized by hepatocytes and excreted into the gallbladder through the capillary duct and bile duct, and enters the duodenum with bile to help digest food. . 95% of bile acid is reabsorbed in the terminal ileum, returns to the liver via the portal vein, and is taken up by hepatocytes for reuse. It mainly exists in the form of protein binding in serum, and the total amou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543
CPCG01N33/54313
Inventor 冯志军
Owner 北京美德美康生物技术有限公司
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