Double-particle compounded C-reactive protein detection kit

A technology for detection kits and reactive proteins, which is applied in the field of medical immunology in vitro diagnosis, to achieve the effects of avoiding non-specific reactions, improving purity and potency, and high sensitivity

Inactive Publication Date: 2012-12-05
上海睿康生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, immunoturbidimetry is the most common method, but the highest detection limit of immunoturbidimetry can only reach 8mg / l, and the linearity can only reach 150mg / l, which has limitations in clinical application.

Method used

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  • Double-particle compounded C-reactive protein detection kit

Examples

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Comparison scheme
Effect test

preparation example Construction

[0016] The preparation method of the reagent R2 is as follows: the polystyrene latex particles of 156nm and the polystyrene latex particles of 61nm are mixed according to the ratio of 1:5, the goat anti-human CRP polyclonal antibody and the mixed polystyrene latex are mixed with The mass ratio of 50:100 was mixed, and the buffer was 0.02M phosphate buffer. After mixing the two, they were adsorbed at 37°C for 8 hours, and then dialyzed to remove the unlinked antibody. Glycine, blocked for 2 hours, centrifuged to remove the supernatant, diluted to 0.18% with PH7.4 phosphate buffer 45mmol / l, disodium edetate 10.2mmol / l, 0.05% skimmed milk powder, and 0.9% NaN3.

[0017] The preparation method of the calibration product is as follows: add BAS 0.1% and NaN3 0.9% to the processed bovine serum to obtain a calibration product dilution, dissolve the recombinant CRP protein in the prepared solution similar to the human serum matrix, and prepare standards of different concentrations Tast...

Embodiment 1

[0019] Embodiment 1: Correlation test, with reference to figure 1 : Use the CRP latex enhanced reagent of this method invention reagent and contrast reagent A company, adopt automatic 7170 full-automatic biochemical analyzers to measure 85 parts of human serum simultaneously by respective parameter, measured value is carried out correlation analysis. Measure according to the parameters in the above-mentioned "CRP assay method", and the X and Y axes are all measured values ​​(the content of CRP in mg / L).

[0020] Depend on figure 1 The results show that the correlation between the two reagents is R 2 =0.987, and the regression equation is y=0.983x+4.096. The results show that this reagent has a good correlation with the imported reagent in the determination of patient serum, and has good specificity and accuracy.

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Abstract

The invention provides a double-particle compounded C-reactive protein detection kit and relates to the field of in vitro diagnosis of medical immunology. The kit consists of three parts, namely an R1 reagent, an R2 reagent and a calibrator, wherein the R1 reagent is a phosphate buffer system and comprises the following components in mass ratio: 30-80mmol / l of phosphate buffer with the pH value of 7.0-7.6, 60-120mm / l of polyethylene glycol 6000-10000 and 10-20 mmol / L of disodium edta; the R2 reagent is an antibody sensitized grain suspension and comprises compounded CRP (C-Reactive Protein) antibody particles, phosphate buffer with the pH value of 7.0-7.6, disodium edta and the like; and the calibrator is a bovine serum substrate and comprises 0.2-2.2% of sodium azide, 1-10% of tween-20 and 1-3% of bovine serum albumin. The kit provided by the invention adopts an antibody coated with compounded latex particles, is high in sensitivity and can be used for detecting 0.2mg / l of samples and improving the linear value.

Description

Technical field: [0001] The invention relates to the field of medical immune in vitro diagnosis, in particular to a double-particle composite C-reactive protein detection kit. Background technique: [0002] C-reactive protein, named after Tillet and Francis found in the serum of patients with acute lobar pneumonia in 1930 that it can precipitate and react with pneumococcal C polysaccharide in the presence of calcium ions, is an important acute-phase reactive protein in humans, and its concentration in the acute phase can vary. Increased by thousands of times, the half-life of CRP in the circulation is 19 hours. Human CRP is produced by the liver and consists of five identical subunits that rely on non-covalent bonds to form a cyclic pentamer. This characteristic structure classifies it into pentaxins (a group of calcium-binding proteins with immune defense properties). protein) family. CRP also exists in lower animals, such as horseshoe crabs, river mussels, etc., but it d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/536
Inventor 房君江李伟奇张秀文林清玉
Owner 上海睿康生物科技有限公司
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