Kit for quantitatively detecting CEA (Carcino-Embryonic Antigen) as well as preparation method and application method of kit

A quantitative detection and kit technology, applied in in vitro clinical testing and nanometer field, to achieve the effects of rapid response, high sensitivity and wide linear range

Inactive Publication Date: 2014-02-12
INST OF ELECTRONICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to propose a new kit for quantitative detection of CEA and its preparation and use methods, so as to solve the problem that the existing technology cannot detect CEA simply, quickly, sensitively and at low cost.

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  • Kit for quantitatively detecting CEA (Carcino-Embryonic Antigen) as well as preparation method and application method of kit
  • Kit for quantitatively detecting CEA (Carcino-Embryonic Antigen) as well as preparation method and application method of kit

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Embodiment 1: Kit

[0031] This embodiment is a CEA quantitative detection kit, which is based on biofunctionalized magnetic nanocomposite particle technology, and has high sensitivity and rapid response. The so-called kit is a set product including multiple reagents, and the various reagents in the kit are used together to complete specific functions.

[0032] According to the present invention, the kit includes at least the following reagents: biofunctionalized magnetic nanocomposite particles coupled with CEA antibody, enzyme-labeled CEA antibody and chemiluminescent substrate solution. The biofunctionalized magnetic nanocomposite particles coupled with CEA antibody are used to capture CEA antigen and separate CEA from other media under the action of an external magnetic field; the enzyme-labeled CEA antibody is used to combine with CEA and biofunctionalized magnetic nanocomposite particles A sandwich structure is formed and the chemiluminescent substrate is catalyz...

Embodiment 2

[0040] Embodiment 2: the preparation method of kit

[0041] The preparation method of the kit includes the steps of preparing each reagent in the kit. In the present invention, it mainly lies in the preparation method of biologically functionalized magnetic nanocomposite particles, which is also the innovation and key of the present invention. Biologically functionalized magnetic nanocomposite particles are microspheres composed of superparamagnetic nanoparticles, and biological antibodies are coupled to the microspheres to form biofunctionalized magnetic nanocomposite particles.

[0042] In the first step, the biotinylated CEA antibody is added to the magnetic beads, and shaken for a period of time,

[0043]The two are coupled to form biofunctionalized magnetic nanocomposite particles; in the second step, BSA (bovine serum albumin) is added to the magnetic beads and the shaking reaction is performed again to block the unreacted sites on the magnetic beads; in the third step, ...

Embodiment 3

[0049] Embodiment 3: the method for using kit to detect CEA

[0050] 1. Pretreatment of kits and samples.

[0051] In this embodiment, the kit was placed at room temperature (18-26° C.) to equilibrate for 10-15 minutes; 100 μL of the concentrated washing solution was diluted with deionized water at 1:100 for later use; the samples to be tested (blood, pleural volume liquid, sputum, saliva, semen, urine, ascites), etc., take the supernatant, and put it at room temperature for 10-15 minutes to equilibrate.

[0052] 2. Using the kit to detect CEA on the sample.

[0053] Step 1: This step is the detection process of the calibrator, and the calibrator is used for calibration. This step consists of the following three steps:

[0054] In the first step, the biofunctionalized magnetic nanocomposite particles are used in this step to capture the CEA in the solution. In this embodiment, 50 μL of biofunctionalized magnetic nanocomposite particles and 50 μL of calibrator are mixed, ta...

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Abstract

The invention discloses a kit for quantitatively detecting a CEA (Carcino-Embryonic Antigen). The kit comprises following reagents: a biological functional magnetic nano composite particle coupled with a CEA antibody, an enzyme-labeled CEA antibody and a chemiluminiscence substrate, wherein the biological functional magnetic nano composite particle coupled with the CEA antibody is formed by compounding a nano-grade magnetic nucleus with a high molecular material containing various functional groups. According to the kit for quantitatively detecting the CEA, a nano magnetic bead separation technology, a chemiluminiscence technology and an enzyme immunoassay technology are combined together; the kit has the advantages of high sensitivity, wide linearity range, high specificity, rapidness in reaction and the like.

Description

technical field [0001] The invention belongs to the fields of in vitro clinical testing and nanotechnology, and in particular relates to a kit for quantitatively detecting carcinoembryonic antigen (CEA), a preparation method thereof and a method for detecting CEA using the kit. Background technique [0002] Cacinoembryonic antigen (CEA) is a highly glycosylated cell surface glycoprotein with a molecular weight of 180-220kDa. CEA is a broad-spectrum tumor marker, which can be expressed at a high level in colorectal cancer, pancreatic cancer, gastric cancer, lung cancer, breast cancer and other diseases. Detecting the content of CEA in body fluids (serum, pleural effusion, sputum, saliva, semen, urine, ascites, etc.) has important clinical significance for tumor diagnosis, prognosis, and postoperative observation. The reference value of normal serum CEA is 5ng / mL. [0003] In 1969, radioimmunoassay was first used in the detection of CEA. Although radioimmunoassay is the mos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/544G01N33/545G01N33/552
CPCG01N33/57473G01N33/535G01N33/54346
Inventor 曲书雪蔡新霞刘军涛黄一清王彬
Owner INST OF ELECTRONICS CHINESE ACAD OF SCI
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