Kit for detecting C-reactive protein through fluorescent immunochromatography

A technology of fluorescence immunochromatography and detection kits, applied in biological testing, measuring devices, analytical materials, etc., can solve the problems of narrow linearity, low sensitivity, unfavorable for high-throughput automatic detection, etc., and achieve the effect of reducing errors

Pending Publication Date: 2015-12-16
NINGBO RUI BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This technique has low sensitivity, narrow linearity, poor spe...

Method used

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  • Kit for detecting C-reactive protein through fluorescent immunochromatography
  • Kit for detecting C-reactive protein through fluorescent immunochromatography
  • Kit for detecting C-reactive protein through fluorescent immunochromatography

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] AlexaFluor660 from Lifetechnologies was used for fluorescein, which was dissolved in DMSO and then adjusted to a concentration of 10 mg / ml as a fluorescein storage solution. The fluorescein was activated by EDC / NHS cross-linking method (the molar ratio of fluorescein to the total amount of chemical cross-linking agent was 1:0.14). The fluorescein storage solution was mixed with EDC and NHS, and 50 mM pH6.0 MES buffer was used as the reaction medium, and incubated at 37° C. for 60 minutes to obtain activated fluorescein.

[0047] Take 1 mg of activated fluorescein and add 20 mg of mouse anti-human CRP monoclonal antibody I (T1) to incubate at 37°C for 60 minutes. After that, free fluorescein was dialyzed with 50 mM Tris-HCl buffer pH7.4, and the dialysate was changed every 6 hours for a total of three times. Collect the liquid in the dialysis bag, use Tris-HCl 50mM / L, 1% BSA, 0.6% Triton X-100, 0.9% NaCl, 0.05% NaN after ultrafiltration and concentration 3 Fluorescein ...

Embodiment 2

[0057] AlexaFluor 660 from Lifetechnologies was used for fluorescein, which was dissolved in DMSO and then adjusted to a concentration of 5 mg / ml as a fluorescein storage solution. The fluorescein was activated by EDC / NHS cross-linking method (the molar ratio of fluorescein to the total amount of chemical cross-linking agent was 1:0.14). The luciferin storage solution was mixed with EDC and NHS, and the activated luciferin was obtained by incubating at 36° C. for 10 minutes with 50 mM pH6.2 MES buffer as the reaction medium.

[0058] Take 1 mg of activated fluorescein and add 5 mg of mouse anti-human CRP monoclonal antibody I (T1) to incubate at 36° C. for 10 minutes. After that, free fluorescein was dialyzed with 50 mM Tris-HCl buffer pH7.4, and the dialysate was changed every 6 hours for a total of three times. Collect the liquid in the dialysis bag, use Tris-HCl 5mM / L, 2.3% BSA, 0.1% Triton X-100, 0.9% NaCl, 0.43% NaN after ultrafiltration and concentration 3 Fluorescein ...

Embodiment 3

[0066] The fluorescein was AlexaFluor 660 from Lifetechnologies, which was dissolved in DMSO and then adjusted to a concentration of 7 mg / ml as a fluorescein storage solution. The fluorescein was activated by EDC / NHS cross-linking method (the molar ratio of fluorescein to the total amount of chemical cross-linking agent was 1:0.14). The luciferin stock solution was mixed with EDC and NHS, and 50 mM pH6.4 MES buffer was used as the reaction medium, and incubated at 38° C. for 120 minutes to obtain activated luciferin.

[0067] Take 1 mg of activated fluorescein and add 50 mg of mouse anti-human CRP monoclonal antibody I (T1) to incubate at 38° C. for 120 minutes. After that, free fluorescein was dialyzed with 50 mM Tris-HCl buffer pH7.4, and the dialysate was changed every 6 hours for a total of three times. Collect the liquid in the dialysis bag, use Tris-HCl 200mM / L, 1.7% BSA, 2% Triton X-100, 0.9% NaCl, 0.85% NaN after ultrafiltration and concentration 3 Fluorescein Antibo...

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Abstract

The invention discloses a kit for detecting C-reactive protein through fluorescent immunochromatography. In the kit, a mouse anti-human CRP (C-reactive protein) monoclonal antibody with a fluorescent label and a rabbit IgG (immunoglobulin G) with a fluorescent label are made by subjecting the mouse anti-human CRP monoclonal antibody and the rabbit IgG to fluorescein labeling; in labeling the mouse anti-human CRP monoclonal antibody and the rabbit IgG with fluorescein, the mouse anti-human CRP monoclonal antibody and the rabbit IgG are added into activated fluorescein storage liquid; the concentration of fluorescein in the fluorescein storage liquid is 5-10mg/mL. The kit is high in sensitivity and good in detection stability, enables reduction in nonspecific binding, has a wider linear range, takes only 3 minutes for detection and enables great improvement in diagnostic efficiency.

Description

technical field [0001] The invention relates to a C-reactive protein detection kit, in particular to a C-reactive protein fluorescent immunochromatography detection kit. Background technique [0002] Serum CRP level is a sensitive and objective indicator of bacterial infection. During bacterial infection, the level of serum CRP can be moderately or significantly increased, and the positive rate can reach more than 90%. However, during viral infection, the level of CRP is mostly normal or slightly elevated, so it can help to distinguish bacterial infection from non-bacterial infection diagnosis. In addition, the quantitative measurement of CRP levels in cerebrospinal fluid and pleural effusion can also be of certain significance in the differential diagnosis of meningitis and pleurisy; not only that, but also the level of CRP has a certain relationship with the scope and severity of infection. In addition, serum CRP levels can also be used to predict the severity of infecti...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577G01N33/558
CPCG01N33/68G01N33/558G01N33/577G01N2333/4737
Inventor 周广亮张闻周海滨王建飞
Owner NINGBO RUI BIO TECH
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