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A hypersensitive c-reactive protein detection kit based on liposome signal amplification

A detection kit and signal amplification technology, applied in the field of CRP detection kits, to achieve the effect of expanding the detection linear range, ensuring antibody stability, and enhancing natural properties

Active Publication Date: 2015-12-30
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

However, there is no report on the use of liposomes as a signal amplification system for immunoturbidimetry in China at present.

Method used

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  • A hypersensitive c-reactive protein detection kit based on liposome signal amplification
  • A hypersensitive c-reactive protein detection kit based on liposome signal amplification
  • A hypersensitive c-reactive protein detection kit based on liposome signal amplification

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] This embodiment relates to a detection kit for hypersensitive C-reactive protein (hs-CRP), which is prepared as follows:

[0038] Reagent R1:

[0039] Glycine buffer

pH6.0~6.5 25mM

NaCl

200mM

Tween20

0.9ml / L

EDTA

100mM

Dextran 6000

300mM

NaN 3

1‰

[0040] Various ingredients can be added sequentially at room temperature, sealed and stored for later use.

[0041] Reagent R2: Dissolve EDC in purified water to prepare a 5mg / ml solution, and equilibrate at room temperature half an hour before preparation. Mix surface carboxylated liposomes with a diameter of 180-235nm (phosphatidylcholine: DPPC=1:1) and mouse anti-human CRP monoclonal antibody at a mass ratio of 10:1.5, add 0.02M, pH5 .0~6.0 phosphate buffer solution, DPPC liposome concentration is 0.1~1.5% (volume ratio), mix, incubate at room temperature for half an hour, then add 5mg / ml EDC solution (containing glutaraldehyde), make EDC react ...

Embodiment 2

[0046] The hs-CRP detection kit described in this example is applicable to various types of automatic biochemical analyzers, and Hitachi 7170 automatic biochemical analyzer is used as an example here.

[0047] Analysis method: two-point endpoint method, that is, the dosage of reagent R1 and reagent R2 is 140ul and 140ul respectively, and the sample volume is 2ul. Adopt this reagent and above-mentioned assay method, adopt the curve (as figure 1 Shown), each point represents a content of the reference calibrator, wherein the X-axis represents the hs-CRP content (mg / ml); Y-axis represents the absorbance.

Embodiment 3

[0048] Example 3: Linear range

[0049] The high concentration sample (320.62mg / ml) of hs-CRP close to the upper limit of the linear range was diluted with hs-CRP diluent by 1 / 2, 1 / 10, 1 / 15, 1 / 30, 1 / 60 and Low-value sample (0.11mg / ml), altogether 7 solutions of different concentrations, detect each concentration with the method described in embodiment 2, every concentration repeats measurement 3 times, the mean value of measured concentration and theoretical concentration are carried out linear regression analysis, The calculated regression equation is y=1.0218x+0.6204, and the correlation coefficient is R 2 =0.9996, showing that the correlation of the kit of the present invention is better in the linear range of 0.1mg / ml~400mg / ml, see figure 2 .

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Abstract

The invention relates to a liposome signal amplification-based hypersensitive C-reactive protein detection kit. The kit comprises an R1 reagent, an R2 reagent and a calibration substance; the R1 reagent is a glycine buffer system; the R2 reagent is a CRP antibody sensitized particle suspension; and the calibration substance is a bovine serum matrix containing 6 CRP recombinant proteins with different concentrations, same solvate and same content. The kit is a kit using latex enhanced immunoturbidimetry, and a CRP antibody is bonded to the surface of liposome through a chemical bond. Compared with products sold in the present market, the hypersensitive C-reactive protein detection kit has the advantages of high sensitivity and wide linearity.

Description

technical field [0001] The invention relates to a hypersensitive C-reactive protein detection kit based on liposome signal amplification, in particular to a CRP detection kit using liposome as a signal amplification carrier. Background technique [0002] Hypersensitive C-reactive protein (hypersensitive3C-reactiveprotein, hs-CRP) is a kind of C-reactive protein in plasma. C-reactive protein is a non-specific marker of the acute phase of systemic inflammatory response synthesized by the liver and is one of the most powerful predictors of cardiovascular event risk. hs-CRP hypersensitive C-reactive protein is a sensitive indicator to distinguish low-level inflammatory states, and serum hs-CRP levels are closely related to the occurrence, severity and prognosis of atherosclerosis and acute cerebral infarction (ACI). Clinical laboratories use ultra-sensitive detection technology to accurately detect low-concentration C-reactive protein, which improves the sensitivity and accurac...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/531
CPCG01N33/92G01N33/96
Inventor 李伟奇赵小凤房君江张秀文林清玉
Owner 上海睿康生物科技有限公司