A test strip for detection of ev71 virus iga antibody

An antibody detection and test strip technology, applied in the field of medical testing, can solve the problems of expensive fluorescence detectors and cannot be promoted, and achieve the effect of avoiding spread and shortening the course of disease.

Active Publication Date: 2020-10-09
GUANGZHOU WOMEN AND CHILDRENS MEDICAL CENTER
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because RT-PCR fluorescence detection technology requires strictly certified laboratory conditions, expensive fluorescence detectors, and professionally trained technicians, it cannot be promoted in grassroots units such as community clinics and kindergartens.
The serum IgM antibody detection method is mainly to detect the IgM antibody in the patient's serum to judge the EV71 virus infection. The main technical methods include enzyme-linked immunosorbent technology and colloidal gold rapid chromatography technology, and there are certain false positives and false negatives

Method used

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  • A test strip for detection of ev71 virus iga antibody
  • A test strip for detection of ev71 virus iga antibody
  • A test strip for detection of ev71 virus iga antibody

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Embodiment 1: Preparation of diagnostic test strip coating

[0026] The amino acid sequence of the EV71 virus VP2 protein antigen polypeptide is shown in SEQ ID NO.1. The invention adopts the bifunctional agent maleimido succinimidyl benzoate (MBS) to couple the BSA protein to the EV71 virus VP2 protein antigen polypeptide, and the coupler can be used for coating diagnostic test strips. 3 mg of MBS was weighed and dissolved in dimethylamide (DMF) to prepare an MBS solution with a concentration of 10 mM. Dissolve 3 mg of BSA in 1 ml of 10 mM PBS (pH 7.2), add 100 ul of 10 mM MBS solution, and stir magnetically at room temperature for 10-30 min to obtain a BSA / MBS solution. Purify the BSA / MBS solution with a sepharose G25 chromatography column, use 50 mM PBS (pH6.0) to equilibrate the column and elute the protein, and collect the BSA / MBS protein. Dissolve 5mg of EV71 virus VP2 protein antigen polypeptide (as shown in SEQ ID NO.1) in 100uL dimethylamide (DMF), then add 1...

Embodiment 2

[0027] Example 2: Preparation of goat anti-human IgA antibody quantum dot labeling pad

[0028] Draw 30ul of water-soluble quantum dots (surface carboxyl modification), centrifuge at 6000rpm for 8min, add 100ul of 10mM PBS (pH 7.2) to dissolve. Add mass fraction 2% N-hydroxyl sulfosuccinimide and mass fraction 2% 1-(3-dimethylaminopropyl)-3-ethyl carbodiamine hydrochloride of 30ul volume in the quantum dot solution, Add 1000 ul of 5 mg / ml goat anti-human IgA antibody solution and stir at room temperature for 4 hours. After the reaction was complete, centrifuge at 6000 rpm for 8 min, and add 1% BSA solution with a mass fraction of the precipitate to block for 2 hours. Centrifuge at 6000rpm for 8min, add 10mM PBS (pH 7.2) to dissolve the precipitate, and obtain goat anti-human IgA antibody quantum dot microspheres, which are stored at 4°C for later use. Spray the goat anti-human IgA antibody quantum dot microsphere marker on the glass fiber membrane, the dilution parameter is ...

Embodiment 3

[0029] Example 3: Nitrocellulose membrane coating

[0030] Dilute the human IgA antibody and EV71 virus VP2 protein antigen polypeptide-BSA protein with coating solution, wherein the concentration of human IgA antibody is 1.5 mg / ml, and the EV71 virus VP2 protein antigen polypeptide-BSA protein concentration is 1 mg / ml. The two are evenly sprayed onto the NC membrane, wherein the human IgA antibody is sprayed onto the C (control line) position on the NC membrane, and the EV71 virus VP2 antigen polypeptide-BSA protein is sprayed onto the T (test line) position on the NC membrane. Place the sprayed NC film at a relative humidity of 20% and a temperature of 20-35°C for drying treatment for more than 12 hours, and seal the dried NC film for later use.

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Abstract

The invention discloses a test strip for detecting EV71 virus IgA antibody. It comprises a sample pad, a marker pad, a nitrocellulose membrane, an absorbent paper and a backboard, and is characterized in that, the nitrocellulose membrane is provided with a detection line containing an antigen and a control line of human IgA antibody, and the marker pad contains Anti-human IgA antibody quantum microsphere marker; the antigen includes EV71 virus VP2 protein antigen polypeptide, and its amino acid sequence is shown in the sequence table SEQ ID NO.1. The invention is more suitable for kindergartens to screen hand, foot and mouth virus, and can screen suspected infected children without professional medical personnel, and can avoid the spread of EV71 virus in kindergartens. The present invention is also suitable for family diagnosis. Parents can know the condition of their children in advance, and send them to the hospital for diagnosis and treatment as soon as possible, so as to shorten the course of the disease.

Description

technical field [0001] The invention belongs to the field of medical inspection, and specifically discloses a test strip for detecting IgA antibody of EV71 virus. Background technique [0002] The EV71 virus is a member of the enterovirus, and generally infects children under the age of 6. Common symptoms mainly include hair circles, loss of appetite, sore throat, oral ulcers, blisters on the hands and feet, etc., and severe cases can lead to aseptic meningitis, cerebral palsy, etc. Serious central nervous system complications such as inflammation, poliomyelitis-like paralysis, and neurogenic pulmonary edema, and death. The incidence of EV71 virus has a certain seasonality. Generally, April to July is the high incidence period of virus infection, and there are sporadic infection cases in other months. It is the virus with the largest number of deaths among children. At present, there is no special drug to treat EV71 virus. Clinically, it is basically symptomatic treatment. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/533G01N33/569
CPCG01N33/533G01N33/56983
Inventor 王长兵朱冰李英华邝璐韩宁陈翊许甜甜林正方
Owner GUANGZHOU WOMEN AND CHILDRENS MEDICAL CENTER
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