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Fluorescent immunochromatography method for whole quantitative detection of C-reactive protein and reagent kit thereof

A technology of fluorescent immunochromatography and reactive protein, which is applied in the field of fluorescent immunochromatographic method and its kit for quantitative detection of C-reactive protein in the whole process, which can solve the problems of low sensitivity and narrow detection range of C-reactive protein, and achieve the goal of expanding Effects of improved detection range, improved quantitative accuracy, and improved sensitivity

Active Publication Date: 2012-07-04
SHENZHEN KANGMEI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] The technical problem to be solved in the present invention is to provide a fluorescent immunochromatographic method and its kit for the full quantitative detection of C-reactive protein in view of the shortcomings of low sensitivity and narrow detection range in the prior art for detecting C-reactive protein

Method used

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  • Fluorescent immunochromatography method for whole quantitative detection of C-reactive protein and reagent kit thereof
  • Fluorescent immunochromatography method for whole quantitative detection of C-reactive protein and reagent kit thereof
  • Fluorescent immunochromatography method for whole quantitative detection of C-reactive protein and reagent kit thereof

Examples

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Effect test

Embodiment 1

[0072] Example 1: Quantitative detection of CRP by modifying antibodies to quantum dots by chemical cross-linking and using direct pre-wetting immunochromatography

[0073] (1) Modification of quantum dots and antibodies

[0074] Quantum dots with an emission wavelength of 650nm were mixed with 1mg / mL CRP monoclonal antibody, and mixed with freshly prepared N-hydroxysuccinimide (NHS, 1mg / mL) and carbodiimide hydrochloride (EDC, 1mg / mL) under the action of room temperature for 4 to 5 hours, adding 1mol / L glycine to block, and using a chromatographic column or chromatographic column to separate and purify to obtain CRP monoclonal antibody modified quantum dots. Similarly, rabbit IgG-modified quantum dots were obtained. The fluorescence emission wavelength of the CRP antibody-modified quantum dot is 650nm, and the fluorescence emission wavelength of the rabbit IgG-modified quantum dot is 570nm.

[0075] (2) Construction of the kit

[0076] Mix the two kinds of quantum dot mar...

Embodiment 2

[0088] Example 2: Quantitative detection of CRP by modifying antibody to quantum dots with biotin-avidin system and using indirect pre-wetting immunochromatography

[0089] (1) Modification of quantum dots and antibodies

[0090] First, 1 mL of CRP monoclonal antibody (1 mg / mL) was fully dialyzed with pH 9.0 sodium bicarbonate buffer, and 20-120 μl of N-hydroxysuccinimide biotin ester freshly prepared in dimethyl sulfoxide (DMSO) was added (NHSB, 1mg / mL), react at room temperature in the dark for 4h. Add 1mol / L NH 4 Cl, shaking reaction at room temperature for 10 minutes, then put the solution in a dialysis bag, dialysis and purification overnight, and concentrate with an ultrafiltration centrifuge tube to prepare the required concentration, and the obtained biotinylated CRP monoclonal antibody.

[0091] Streptavidin-modified quantum dots with an emission wavelength of 900nm and biotinylated CRP monoclonal antibody were mixed and reacted at a ratio of 1:3-1:12 for 30-60 minu...

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Abstract

The invention discloses a fluorescent immunochromatography method for whole quantitative detection of C-reactive protein and a reagent kit thereof. The fluorescent immunochromatography method for the whole quantitative detection of the C-reactive protein (CRP) utilizes excellent fluorescent characteristics of quantum dots, and combines double-color marking technology and immunochromatography technology to achieve fluorescent quantitative detection on the basis of optimizing each constituent elements of test paper. Compared with a conventional colloidal gold immunochromatography method, the fluorescent immunochromatography method for the whole quantitative detection of the CRP has the advantages of being good in stability, low in non-specificity, high in flexibility, wide in linear range and accurate in quantifying. The reagent kit of the fluorescent immunochromatography method can perform the whole quantifying and can simultaneously predict and evaluate infectious diseases, antibiotic effects and cardiovascular and cerebrovascular diseases. The fluorescent immunochromatography method for the whole quantitative detection of the CRP and the reagent kit of the fluorescent immunochromatography method are suitable for various-level hospitals, and particularly contribute to wide popularization in basic-level hospitals and clinics.

Description

technical field [0001] The invention relates to the field of medical testing, in particular to a fluorescent immunochromatographic method for quantitatively detecting C-reactive protein throughout the whole process and a kit thereof. Background technique [0002] C-reactive protein (C-reactive protein, CRP) is an acute phase reaction protein synthesized by the liver, which is non-covalently linked by 5 identical non-glycosylated subunits, and the relative molecular mass of each subunit 23017, consisting of 206 amino acid residues. It has a variety of biological activities and is considered one of the most sensitive indicators of inflammation. CRP not only has important clinical application prospects for infectious diseases, gestational diabetes course detection and prognosis evaluation, antibiotic efficacy, etc., but also is one of the most powerful predictors of cardiovascular diseases. [0003] CRP contains only a small amount in a healthy human body, and its concentrati...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N21/64
Inventor 王秀利
Owner SHENZHEN KANGMEI BIOTECH
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