CN1904615A

CN1904615A - Influenza A,B virus antigen colloidal gold combined detection test paper

Info

Publication number: CN1904615A
Application number: CN 200510012268
Authority: CN
Inventors: 刘明
Original assignee: BEIJING ZHUANGDI HAOHE BIOMEDICINE SCIENCE AND TECHNOLOGY Co Ltd
Current assignee: BEIJING ZHUANGDI HAOHE BIOMEDICINE SCIENCE AND TECHNOLOGY Co Ltd
Priority date: 2005-07-28
Filing date: 2005-07-28
Publication date: 2007-01-31

Abstract

The invention supplies a rapid testing tape that could simultaneously test A, B virus antigen. It covers FluA-McAb, FluB-McAb and IgG on NC film, compounds FluA-McAb, FluB-McAb marked by colloidal gold, simultaneously tests A, B virus antigen in tested sample by using film chromatography double antibody cream filling method. The invention could decrease cockamamie process, and could gain result in 10-15 minutes. It is suitable to hospital and the research for large scale epidemiology to handle sporadic affair.

Description

A kind of influenza A, B virus antigen colloidal gold combined detection test paper

Technical field

The invention belongs to field of biological detection, be specifically related to a kind of influenza A, B viral antigen test strip and application thereof.

Background technology

The detection of influenza A, B virus at present with the antibody test of tissue culture, paired sera as " goldstandard ", but this method complex operation complexity, time is long, to the timely discovery of influenza, in time forecast and prevention does not have directive function, but the science of heredity of analyzing virus population and antigenicity are changed and find that new virus still has vital role.Influenza fast detecting method commonly used has: directly, indirect fluorescent method (IFA/DFA), enzyme immunoassay (EIA), enzyme assay method (neuraminic acid enzymatic determination), APAAP bridging enzyme mark detection method, nucleic acid amplification (PCR), but all there are and need expensive instrument, complicated operation, time length, need problems such as professional, can not detect two kinds of antigens of influenza A, B virus simultaneously.

Summary of the invention

The purpose of this invention is to provide quickly and easily a kind of, can detect the test strips of influenza A, two kinds of viral antigens of influenza B simultaneously, the inventor is by wrapping simultaneously on nitrocellulose membrane (NC film) by influenza A monoclonal antibody FluA-McAb, influenza B monoclonal antibody FluB-McAb and three bands of anti-mouse IgG, and two kinds of antibody of FluA-McAb, FluB-McAb of application association colloid gold mark, realize on a reagent strip, detecting simultaneously the purpose of two kinds of antigens, finished the present invention.Overcome the prior art complicated operation, loaded down with trivial details, the time long, need instrument, can not detect the deficiency of two kinds of antigens simultaneously.

The invention provides a kind of test strip, it is characterized in that, it comprises:

(1) wraps simultaneously by the nitrocellulose membrane of influenza A monoclonal antibody FluA-McAb, influenza B monoclonal antibody FluB-McAb and three bands of the anti-mouse IgG of Quality Control;

(2) contain the glass fibre membrane of colloid gold label FluA-McAb and colloid gold label FluB-McAb.

Preferred test strips of the present invention, it also comprises:

(1) reaction holder;

(2) adsorptive pads;

(3) golden labeling antibody diaphragm has absorbent function simultaneously.

Wherein react the preferred PVC plate of holder; The preferred filter paper for oil of adsorptive pads; Gold labeling antibody diaphragm is preferably used polyester film, glass fibre or filter paper fibre.

The invention also discloses the application of described test strips in detecting influenza A, two kinds of viral antigens of influenza B.

Technical scheme of the present invention is: the influenza A, B monoclonal antibody (FluA, B-McAb) and the anti-mouse IgG difference solid phase (NC film) on nitrocellulose membrane that adopt purifying, FluA, the B-McAb of association colloid gold mark use rete and analyse the principle of double antibody sandwich method and detect influenza A, B virus in the nasopharyngeal secretions.

The invention has the beneficial effects as follows: a sample, a test strips can detect two kinds of viral antigens of influenza A, B simultaneously.Save the formality of secondary detection, easily and fast, simple and direct, do not need special instruments and equipment, do not need professional training, the result is clear easily to be distinguished, simple to operate, is easy to promote, and is fit to basic unit, the mass detection that is suitable for accident is fit to stream and transfers, and the auxiliary detection effect is played in the diagnosis of influenza.

Description of drawings

Fig. 1: the front schematic view of A test strips of the present invention;

The side schematic view of B test strips of the present invention;

1: adsorptive pads;

2: (A: bag is by the band of FluA-McAb for nitrocellulose membrane; B: bag is by the band of FluB-McAb; C: bag is by the Quality Control band of anti-mouse IgG);

3: the glass fibre membrane that contains colloid gold label FluA-McAb and colloid gold label FluB-McAb;

4: golden labeling antibody diaphragm;

5: the reaction holder.

Fig. 2: testing result synoptic diagram;

Be followed successively by: influenza A, the B positive; The influenza A positive; The influenza B positive; Negative; Invalid.

Embodiment

Embodiment 1: influenza A, B viral antigen combined detection test paper (referring to Fig. 1)

The reaction holder is 6.2cm * 0.4cm PCV plate; Adsorptive pads is the filter paper for oil of 2cm * 0.4cm; 1.8cm the nitrocellulose membrane of * 0.4cm wraps successively by anti-mouse IgG, FluA-McAb, FluB-McAb; The FluA, the B-McAb glass fibre membrane that contain 0.4cm * 0.4cm colloid gold label; Gold labeling antibody diaphragm is the polyester film of 2.7cm * 0.4cm; Influenza A, B viral antigen combined detection test paper have promptly been formed.

Embodiment 2: detection method (referring to Fig. 2)

Leave and take pharynx nasalis secretion, put into the bottle that cell pyrolysis liquid is housed, embodiment 1 test strips " 4 " is located to insert in the bottle (can take out test strips after observation finishes fruit), the liquid in the sample picks up up, 10-15 minute sentence read result:

As contain influenza A or B antigen, then with test strips on the influenza A or the B-McAb of colloid gold label form corresponding compound, up be coated on nitrocellulose membrane on influenza A or B-McAb combine, form red lines, promptly locate to form red stripes at " A " or " B ".

No matter whether contain corresponding antigen, colloid gold label McAb continues upwards to creep and the anti-mouse IgG that is coated on the film forms the red precipitate line, promptly locates to form red stripes at " C ".This line is a nature controlling line, loses efficacy as collaurum, and this line just can not occur, and illustrates that test strips lost efficacy.

2 or 3 red precipitate lines can appear in positive findings, and 1 red precipitate line appears in negative findings, lose efficacy as lines explanation test strips not occurring.

Experimental example 1: clinical detection result

1005 routine influenza sample patient's sample after testing, influenza A, B virus infect simultaneously, A, B, C three bands 1 example occur, account for 0.01%; Influenza A virus infects, and A, C two bands 374 examples occur, accounts for 37.2%; B, C two bands 413 examples appear in influenza B virus infections, account for 41.1%.Compare sensitivity 99.8%, specificity 100% with the present conventional indirect fluorescent method of using.

Claims

1, a kind of test strip is characterized in that, it comprises:

2, according to the described test strips of claim 1, it also comprises:

(1) reaction holder;

(2) adsorptive pads;

(3) golden labeling antibody diaphragm.

3,, wherein react holder and select the PVC plate for use according to the described test strips of claim 2.

4, according to the described test strips of claim 2, wherein adsorptive pads is selected filter paper for oil for use.

5, according to the described test strips of claim 2, wherein golden labeling antibody diaphragm is selected polyester film, glass fibre or filter paper fibre for use.

6, the application of the arbitrary described test strips of claim 1-5 in detecting influenza A, two kinds of viral antigens of influenza B.