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Fluorescence immunoassay test strip used for detecting tumor marker CA72-4 and preparation method of fluorescence immunoassay test strip

A tumor marker, fluorescent immunology technology, which is applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of long detection time, insufficient sensitivity and anti-interference ability, and high detection cost, and achieves easy operation, rapid detection, and timely detection. The effect of detection

Inactive Publication Date: 2015-11-25
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the limitations of the methodology itself, the sensitivity and anti-interference ability of radioimmunoassay detection is seriously insufficient, and it has basically withdrawn from the market
Enzyme-linked immunoassay technology and chemiluminescence technology are the main technologies for clinical detection of CA72-4 at present, but the detection cost is high and the detection time is long

Method used

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  • Fluorescence immunoassay test strip used for detecting tumor marker CA72-4 and preparation method of fluorescence immunoassay test strip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Quantum dot-coupled antibody method:

[0043] 1. Quantum dot pretreatment:

[0044] After adding 0.01MMES (pH6.0) to 100ul quantum dots (excitation wavelength is 365nm, emission wavelength is 620nm), shake the above mixture on the oscillator for 5 seconds, centrifuge at 14000-20000rpm in a centrifuge for 5 minutes, discard clear.

[0045] 2. Quantum dot antibody coupling:

[0046] 1) Reagent preparation:

[0047] 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide (EDC for short), N-hydroxysuccinimide (NHS for short), bovine serum albumin (BSA for short), anti-CA72-4 Monoclonal Antibody CC49.

[0048] 2) Quantum dot antibody coupling:

[0049] According to the molar ratio of n(QDs):n(EDC)=1:4000, the quantum dots were added with 0.01MMES (pH6.0) and reacted in the dark on a rotary mixer for 15 minutes, centrifuged (14000-20000rmp, 2min), Discard the supernatant, then according to the molar ratio n(QDs):n(NHS)=1:2000, add 100ul 0.02MPBS buffer solution (pH7.2)...

Embodiment 2

[0050] Embodiment 2: Processing of Quantum Dot Labeled Binding Pad and Sample Pad

[0051] 1) Quantum dot label binding pad processing method:

[0052] Soak the glass cellulose membrane in ultrapure water for 15 minutes, then soak it in absolute ethanol for 15 minutes, and dry it in a drying oven at 50°C. Put the quantum dot bonded pad after the above conditions on the bonded pad for treatment. (0.02MBST containing 5% sucrose by mass, 2% trehalose by mass, pH 7.4) for 30 minutes, and then dried overnight (16-20 hours) at 37°C in a drying oven.

[0053] 2) Sample pad processing method:

[0054] Soak the glass cellulose membrane in ultrapure water for 15 minutes, then soak it in absolute ethanol for 15 minutes, and then dry it in a drying oven at 50°C; place the sample pad treated under the above conditions in the sample pad treatment solution (contain mass percent 2%NaCl, mass percent 0.2%PVP, mass percent 0.1%Tween-20, mass percent 0.5%BSA, soak in 0.02MBS of pH7.4 after 30 ...

Embodiment 3

[0055] Example 3: Preparation of Quantum Dot Labeled Conjugation Pads

[0056] Use a pipette gun to draw 5ul of the quantum dot-labeled antibody, evenly drop it on the pre-treated binding pad, and place it in a clean plastic bucket with a desiccant and away from light. After it dries, the quantum dot-labeled antibody will be obtained. binding pad.

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Abstract

The invention provides a fluorescence immunoassay test strip used for detecting a tumor marker CA72-4 and a preparation method of the fluorescence immunoassay test strip. The test strip comprises a sample cushion, a quantum dot marker combination pad, a nitrocellulose membrane and a piece of water absorption paper which are sequentially laid on a bottom plate in an overlapped mode. The double-antibody sandwich immunochromatographic method is adopted. A CA72-4 resistant monoclonal antibody CC49 coating provided with a quantum dot marker is arranged on the quantum dot marker combination pad. A detection line and a quality control line are arranged on the nitrocellulose membrane. A CA72-4 monoclonal antibody B72.3 is arranged on the detection line. A goat-anti-mouse IgG is arranged on the quality control line. The preparation method mainly relates to preparation of the quantum dot marker antibody. After immunochromatography is conducted through the test strip, the quantitative result of CA72-4 is obtained by conducting analysis through a fluorescence immunochromatographic chip detection instrument, operation is easy and convenient, bedside timely and rapid detection can be conducted, and stability and flexibility are high.

Description

technical field [0001] The present invention relates to a test strip for rapid quantitative detection of blood samples and a detection method, in particular to a test strip for rapid quantitative detection of carbohydrate antigen 72-4 (abbreviated as CA72-4) in serum or plasma using quantum dot technology. The testing method belongs to the field of tumor marker detection. Background technique [0002] Gastric cancer is a malignant tumor derived from gastric mucosal epithelium, which accounts for the third of all malignant tumors and the first of digestive tract malignant tumors, and the 5-year survival rate of gastric cancer patients after diagnosis is very low. [0003] CA72-4 is a tumor-associated glycoprotein obtained by Coleher et al. from liver metastases of breast cancer in 1981. Its molecular weight is greater than 1,000,000, and it belongs to mucin carcinoid embryonic antigen. Histochemical studies have shown that it exists in 50% of breast cancer and 85%-95% of col...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/533
CPCG01N33/533G01N33/577
Inventor 王侃闫薪宇崔大祥何井华秦伟健
Owner SHANGHAI JIAO TONG UNIV
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