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Kit for determining heart-type fatty acid binding protein in serum or urine by latex enhanced turbidimetric immunoassay

A fatty acid binding, immunoturbidimetric technology, applied in the field of medical laboratory science, can solve the problems of low sensitivity, cumbersome steps, radioactive pollution, etc., achieve high linearity and sensitivity, improve detection efficiency, and improve the effect of accuracy

Active Publication Date: 2012-08-08
BEIJING STRONG BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the known detection methods of cardiac fatty acid binding protein include radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), immunochromatography, etc., but the expected results of clinical application are not ideal, such as radioimmunoassay steps are cumbersome, Reagents are expensive, need to use supporting equipment and have radioactive contamination; immunochromatography is mostly qualitative detection, low sensitivity; enzyme-linked immunosorbent assay has long detection time, complicated operation, poor repeatability, and is not suitable for timely diagnosis of emergency and clinical patients needs

Method used

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  • Kit for determining heart-type fatty acid binding protein in serum or urine by latex enhanced turbidimetric immunoassay
  • Kit for determining heart-type fatty acid binding protein in serum or urine by latex enhanced turbidimetric immunoassay
  • Kit for determining heart-type fatty acid binding protein in serum or urine by latex enhanced turbidimetric immunoassay

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Kit for the determination of cardiac fatty acid binding protein in serum or urine by latex-enhanced immunoturbidimetric method:

[0035] 1. The main components and concentrations of the kit are as follows:

[0036] Reagent R1:

[0037]

[0038]

[0039] Reagent R2:

[0040]

[0041] Sensitized latex particles coated with mouse anti-human H-FABP monoclonal antibody and goat anti-human H-FABP polyclonal antibody Latex particle size: 160nm Latex concentration 0.2%

[0042] H-FABP calibrator:

[0043]

[0044] According to the required H-FABP reference calibrator concentration, 120 μg / L of the corresponding heart-type fatty acid binding protein pure product was added to the above buffer to prepare a 120 μg / L concentration of H-FABP reference calibrator, which was diluted with normal saline when used. Multi-concentration reference calibrators (120 μg / L, 60 μg / L, 30 μg / L, 15 μg / L, 7.5 μg / L).

[0045] Reagent R1 is a colorless transparent liquid, R2 is a milky ...

Embodiment 2

[0057] Kit for the determination of cardiac fatty acid binding protein in serum or urine by latex-enhanced immunoturbidimetric method:

[0058] 1. The main components and concentrations of the kit are as follows:

[0059] Reagent R1:

[0060]

[0061] Reagent R2:

[0062]

[0063]

[0064] Sensitized latex particles coated with mouse anti-human H-FABP monoclonal antibody and rabbit anti-human H-FABP polyclonal antibody Latex particle size: 160nm Latex concentration 0.2%

[0065] H-FABP calibrator:

[0066]

[0067] According to the concentration of H-FABP reference calibrator, add the corresponding pure heart-type fatty acid binding protein to the above buffer respectively, and prepare the concentrations as 100 μg / L, 50 μg / L, 25 μg / L, 12.5 μg / L, 6.25 μg / L of multi-concentration H-FABP reference calibrator.

[0068] Reagent R1 is a colorless transparent liquid, R2 is a milky white liquid, and the calibrator is a light yellow transparent solution.

[0069] 2. K...

Embodiment 3

[0079] Example 3 Performance evaluation of the heart-type fatty acid binding protein kit

[0080] I. Relevance:

[0081] The kit 1 of the present invention prepared according to Example 1 was tested for correlation with the commercially available kit A (purchased from Biocheck, USA) using enzyme-linked immunosorbent assay (ELISA). Detect 50 fresh human sera (including normal and abnormal samples), measure according to their respective determination methods, and perform correlation analysis on the measured values ​​(results see figure 1 , X and Y axes are measured values, unit ng / ml). Correlation coefficient R 2 =0.913, the regression equation is y=0.837x+0.845, the result shows that the reagent of the present invention has a good correlation with ELISA. The correlation experiment data are shown in Table 1:

[0082] The kit 2 of the present invention prepared according to Example 2 and the commercially available imported kit B were tested for correlation. Detect 50 fresh h...

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Abstract

The invention relates to a kit for determining heart-type fatty acid binding protein in serum or urine by latex enhanced turbidimetric immunoassay. Specifically, the kit for determining the heart-type fatty acid binding protein comprises a reagent R1, a reagent R2 and a calibrator, wherein the reagent R1 contains a reaction promoter, an antiseptic, a surfactant, a stabilizing agent, an electrolyte and a buffer; the reagent R2 contains latex particles with binding of anti-heart-type fatty acid binding protein monoclonal antibody and polyclonal antibody, an antiseptic, a surfactant, a stabilizing agent, an electrolyte and a buffer; and the calibrator contains an antiseptic, an electrolyte, a stabilizing agent, a heart-type fatty acid binding protein pure product and a buffer. By the complex coating method of latex particles with the monoclonal antibody and the polyclonal antibody, high sensitivity and wide linear range of the kit are guaranteed. Simultaneously, the kit also has advantages of high accuracy, good repeatability, strong singularity, easy operation and the like, and is applicable to an automatic biochemical analyzer which is commonly used in clinic.

Description

technical field [0001] The invention belongs to the field of medical laboratory science, and relates to an immunoassay kit, in particular, the invention relates to a kit for measuring heart-type fatty acid binding protein in serum or urine. Background technique [0002] Fatty acid binding protein (FABP) is a family of intracellular proteins with a molecular weight of 14-16kD, including at least nine types of liver, small intestine, heart, brain, kidney, skeletal muscle, adipose tissue, ileum, and epidermis. immunological differences. Heart-type fatty acid binding protein (H-FABP) is a soluble protein present in the cytoplasm of cardiomyocytes, with a molecular weight of 15kD, accounting for 4±8% of the protein content in the myocardium, consisting of 132 amino acids, and is acidic (pI=5). Involved in the uptake, transport and metabolism of cellular fatty acids. The ratio of H-FABP in the cytoplasm of cardiomyocytes to blood is 200,000:1, the concentration in skeletal muscl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N21/31
Inventor 陈婧高爱民蔡华雅刘希
Owner BEIJING STRONG BIOTECH INC
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