Reagent kit and method for joint detection of PCT, CRP and IL-6

A technology of IL-6 and reagent kits, applied in measuring devices, instruments, fluorescence/phosphorescence, etc., to achieve high-accuracy results

Inactive Publication Date: 2016-03-09
SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still a lack of joint detection reagents that can quickly and quantitatively detect the above three indicators at the same time

Method used

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  • Reagent kit and method for joint detection of PCT, CRP and IL-6
  • Reagent kit and method for joint detection of PCT, CRP and IL-6
  • Reagent kit and method for joint detection of PCT, CRP and IL-6

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0042] The preparation method of combined detection PCT, CRP and IL-6 kit generally comprises the following steps:

[0043] (1) Preparation of carboxyl-modified fluorescent microsphere-labeled probes

[0044] Using appropriate carboxyl-modified fluorescent microspheres, after activating the carboxyl groups on its surface, PCT, CRP and IL-6 labeled antibodies are respectively directionally connected to the surface of the carboxyl-modified fluorescent microspheres by means of covalent coupling.

[0045] (2) Coating of antibodies at the T-line and C-line in the test area

[0046] Spray the PCT-coated antibody on the T1 line of the coated film test area, spray the CRP coated antibody on the T2 line of the coated film test area, and spray the IL-6 coated film on the T3 line of the coated film test area by using a film spraying instrument. Antibody, spray goat anti-mouse IgG antibody on C line.

[0047] (3) Coating of the labeled probe at the sample pad

[0048] Spray the anti-PC...

Embodiment 1

[0056] Embodiment 1: the preparation of joint detection PCT, CRP and IL-6 kit

[0057] (1) Preparation of fluorescent microsphere-labeled antibody

[0058] With an average diameter of 110nm, carboxyl-modified fluorescent microspheres (purchased from Bangs Laboratories, Inc., catalog number FC02F / 10930), anti-PCT, CRP, IL-6 monoclonal antibodies (purchased from Yunnan University Monoclonal Antibody Engineering The numbers of the technology center are P-03, C-05, and I6-01), and fluorescent microsphere-labeled PCT, CRP, and IL-6-labeled antibodies are prepared according to the following methods:

[0059] Take 15 mg of the above-mentioned carboxyl-modified fluorescent microspheres, wash and centrifuge with MES buffer (0.1M, pH4.7), resuspend with 1ml MES buffer (0.1M, pH4.7), add 1-ethyl-(3 -Dimethylaminopropyl) carbodiimide (EDC) to a final concentration of 5mM, add NHS (N-hydroxysuccinimide) to a final concentration of 10mM, room temperature away from light, react for half an ...

Embodiment 2

[0065] Embodiment 2: Evaluation of combined detection PCT, CRP, IL-6 kit

[0066] (1) Detection sensitivity

[0067] The sensitivity of the combined detection PCT, CRP, IL-6 kit of Example 1 was determined with PCT, CRP, IL-6 standard items as samples to be tested.

[0068] The PCT standard was prepared as a series of concentrations (0, 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 50, 100ng / mL), and the CRP standard was prepared as a series of concentrations (0, 0.01, 0.05, 0.1, 0.5, 1, 5, 10ug / mL), the IL-6 standard was prepared as a series of concentrations (0, 5, 10, 50, 100, 500, 1000, 2000pg / mL), and the combined Detect the PCT, CRP, and IL-6 kits in the sample wells, and use a fluorescence detector to detect the fluorescence intensity. Detection steps: Return the sample to be tested to room temperature (25°C) before detection, and use a precise pipette to take 60 μl of the sample to be tested and drop it vertically and slowly into the sample addition of the combined detection PCT, ...

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Abstract

The invention discloses a method for joint detection of PCT, CRP and IL-6 and a reagent kit for the method. The reagent kit comprises a first enveloping film and a second enveloping film. One end of the first enveloping film is connected with one end of the second enveloping film, and a labeled antibody mixture is enveloped in at least one area in the first enveloping film and comprises a first labeled antibody, a second labeled antibody and a third labeled antibody. The second enveloping film comprises a first area, a second area, a third area and a fourth area which are separated, wherein a first enveloped antibody, a second enveloped antibody and a third enveloped antibody are enveloped in the first area, the second area and the third area respectively, the first enveloped antibody and the first labeled antibody can be in specific binding with a first antigen, the second enveloped antibody and the second labeled antibody can be in specific binding with a second antigen, the third enveloped antibody and the third labeled antibody can be in specific binding with a third antigen, an antiantibody is enveloped in the fourth area, and the antiantibody can be in specific binding with the first labeled antibody, the second labeled antibody and the third labeled antibody. By means of the reagent kit, PCT, CRP and IL-6 can be quickly and accurately detected at the same time.

Description

technical field [0001] The present invention relates to the field of biological detection. Specifically, the present invention relates to a kit capable of simultaneously detecting various biological substances and its application. More specifically, the present invention relates to a kit and a combined detection of procalcitonin and C reaction protein and interleukin-6 methods. Background technique [0002] Acute phase protein (APP) is a kind of specific protein closely related to infectious inflammation. In recent years, a large number of studies have shown that a variety of positive and negative acute phase proteins have a significant correlation with infectious inflammation, and are more accurate and reliable than detection of leukocytes, erythrocyte sedimentation rate, enzyme activity and content changes, etc. inflammatory markers. [0003] C-reactive protein (Creactive protein, CRP) is synthesized by the liver and is a typical acute phase protein. Its content in norma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/533G01N21/64
CPCG01N33/577G01N21/64G01N33/533
Inventor 马岚蒋宇扬吴峰岑瑜
Owner SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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