Kit (Latex-enhanced immunoturbidimetry) for detecting content of glycocholic acid in blood serum

An immunoturbidimetric and latex-enhanced technology, applied in the field of biochemistry, can solve the problems of expensive reagents, long detection time, complicated operation, etc., and achieve the effects of reliable detection results, shortened detection time, and wide linear range.

Active Publication Date: 2013-02-27
CO HEALTH BEIJING LAB
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The steps of radioimmunoassay are cumbersome, the reagents are expensive, and supporting instruments are required and there is radioactive contamination
Enzyme-linked immunosorbent assay has long detection time, complicated operation, poor repeatability, and is not suitable for emergency and clinical patients in time for diagnosis.

Method used

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  • Kit (Latex-enhanced immunoturbidimetry) for detecting content of glycocholic acid in blood serum
  • Kit (Latex-enhanced immunoturbidimetry) for detecting content of glycocholic acid in blood serum
  • Kit (Latex-enhanced immunoturbidimetry) for detecting content of glycocholic acid in blood serum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Kit for the determination of glycocholic acid in serum by latex-enhanced immunoturbidimetric method:

[0035] The main components and concentrations of the kit are as follows:

[0036] Reagent R1:

[0037] Tris buffer 50mmol / L pH7.2

[0038] Sodium azide 0.8g / L

[0039] Bovine serum albumin 1.5g / L

[0040] Triton X-100 2g / L

[0041] Polyethylene glycol 2000 0.5g / L

[0042] Nacl 0.8g / L

[0043] Reagent R2:

[0044] Tris buffer 100mmol / L pH7.5

[0045] Sodium azide 0.5g / L

[0046] Bovine serum albumin 2g / L

[0047] Tween20 1.5g / L

[0048] Ethylenedimethylamine bromide 0.8g / L

[0049] Nacl 1g / L

[0050] Latex particles coated with rabbit anti-human glycocholic acid polyclonal sensitization, latex particle size: 200nm, latex concentration 0.5%

[0051] Glycocholic Acid Calibrator:

[0052] Tris buffer 30mmol / L pH7.0

[0053] Sodium azide 0.4g / L

[0054] Bovine serum albumin 0.5g / L

[0055] Nacl 0.6g / L

[0056] According to the required concentration of glyco...

Embodiment 2

[0067] Kit for the determination of glycocholic acid in serum by latex-enhanced immunoturbidimetric method:

[0068] 1. The main components and concentrations of the kit are as follows:

[0069] Reagent R1:

[0070] Tris buffer 50mmol / L pH8.0

[0071] Sodium azide 1.2g / L

[0072] Bovine serum albumin 2.0g / L

[0073] Span80 2g / L

[0074] Polyethylene glycol 2000 0.5g / L

[0075] Nacl 1.5g / L

[0076] Reagent R2:

[0077] Tris buffer 100mmol / L pH7.5

[0078] Sodium azide 0.5g / L

[0079] Bovine serum albumin 1g / L

[0080] Span80 1.2g / L

[0081] Ethylenedimethylamine bromide 0.8g / L

[0082] Nacl 1g / L

[0083] Latex particles coated with rabbit anti-human glycocholic acid polyclonal sensitization, latex particle size: 200nm, latex concentration 0.5%

[0084] Glycocholic Acid Calibrator:

[0085] Tris buffer 30mmol / L pH7.0

[0086] Sodium azide 0.4g / L

[0087] Bovine serum albumin 0.5g / L

[0088] Nacl 0.6g / L

[0089] According to the required concentration of glycoch...

Embodiment 3

[0099] Embodiment 3: Glycocholic acid kit performance evaluation

[0100] 1. Linear correlation

[0101] The kit of the present invention prepared in Example 1 and the commercially available glycocholic acid ELISA detection kit A were used to simultaneously detect 46 serum samples, and the correlation of the detection results between the kit of the present invention and the commercially available ELISA detection kit was compared. (results see figure 1 , X and Y axes are measured values, unit mg / L. ) correlation coefficient r=0.9946, and the linear regression equation is: y=0.9854x+0.0623, the results show that the inventive reagent has a better correlation with the detection results of the ELISA kit. The experimental data are shown in Table 1, and the regression equation is shown in the appendix figure 1 .

[0102] Table 1

[0103] sample number Kit 1 Kit A sample number Kit 1 Kit A 1 1.68 1.72 26 2.44 2.39 2 2.51 2.59 27 0.24 0....

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Abstract

The invention relates to a kit of latex-enhanced immunoturbidimetry for detecting content of glycocholic acid in blood serum. Specifically, the provided glycocholic acid detection kit comprises a reagent R1, a reagent R2 and a calibrator, wherein the reagent R1 comprises a reaction promotion agent, a preservative, a surface active agent, a stabilizer, an electrolyte and a buffer solution; the reagent R2 comprises latex particles combined with a glycocholic acid antibody, a preservative, a surface active agent, a stabilizer, an electrolyte and a buffer solution; and the calibrator comprises a preservative, an electrolyte, a stabilizer, glycocholic acid pure products and a buffer solution. The kit for detecting the content of the glycocholic acid in the blood serum, disclosed by the invention, ensures the high sensitivity and wide linear range of a kit by utilizing a method of coating the latex particles by utilizing polyclonal antibodies, also has the advantages of high accuracy, good repeatability, strong specificity, simplicity in operation and the like, and can be applied to a clinical general full automatic biochemical analyzer.

Description

technical field [0001] The invention relates to the technical field of biochemistry, in particular to a kit for measuring glycocholic acid (CG) content in human serum by using a latex-enhanced immune turbidimetric method. Background technique [0002] Serum glycocholic acid (CG) is one of the conjugated bile acids in which bile acid and glycine are combined twice. In liver cells, cholesterol is transformed into primary bile acid through extremely complicated enzymatic reactions. Among them are cholic acid (CA) and chenodeoxycholic acid (CD-CA). There are three hydroxyl groups (C3, C7, C12) on the steroid core of cholic acid, and the hydroxyl group at the end of the side chain is combined with glycine through a peptide bond, with a molecular weight of 462U. [0003] The normal metabolic pathway of CG is the entero-hepatic circulation. CG is synthesized by liver cells, discharged into the gallbladder through the capillaries and bile ducts, and enters the duodenum along with b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/536G01N21/82
Inventor 孟琛
Owner CO HEALTH BEIJING LAB
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