CN107966561A - A kind of test strips and kit for detecting HIV antibody - Google Patents
A kind of test strips and kit for detecting HIV antibody Download PDFInfo
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- CN107966561A CN107966561A CN201711190830.5A CN201711190830A CN107966561A CN 107966561 A CN107966561 A CN 107966561A CN 201711190830 A CN201711190830 A CN 201711190830A CN 107966561 A CN107966561 A CN 107966561A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
- G01N33/56988—HIV or HTLV
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Abstract
The present invention relates to a kind of test strips and kit for detecting HIV antibody.Test strips of the present invention include bottom plate and are pasted onto the sample pad overlapped on the bottom plate and successively, label bonding pad, nitrocellulose filter and water absorption pad；Wherein, the sample pad is handled using sample pad treatment fluid, and the sample pad treatment fluid includes：The polyvinylpyrrolidone K 30 that the polysorbas20 and mass fraction that casein that polyethylene glycol that the PBS buffer of pH7.2~7.5, mass fraction are 0.5~1%, mass fraction are 0.5~1%, volume fraction are 0.3~1% are 0.6~1%；The anti-human IgG antibodies of labeled substance markers are coated with the label bonding pad；Detection line and nature controlling line are provided with the nitrocellulose filter, wherein, HIV antigens are coated with the detection line, antiantibody are coated with the nature controlling line, the antiantibody is with reference to the anti-human IgG antibodies.Test strips of the present invention can amplify the weakly positive testing result of HIV antibody so as to reduce false negative, improve the accuracy of detection.
Description
Technical field
The present invention relates to immuno-chromatographic test paper strip field, in particular to a kind of test strips for detecting HIV antibody and
Kit.
Background technology
AIDS, i.e. Immune Deficiency Syndrome (AIDS), are caused by human immunodeficiency virus (HIV)
A kind of infectious disease for seriously endangering human health.From world's report Patient With Aids case in 1981 so far, 30 years short
Between, AIDS has wreaked havoc the life that more than 2,500 ten thousand people are seized in the whole world.The report of The Joint Programme on AIDS publication year, 2010
Year global about 34,000,000 people of AIDS virus HIV carriers, increase by 700,000 in than 2009.Report display, whole world Chinese mugwort in 2010
Grow sick virus carrier and increase about 2,700,000 newly, 15% was reduced than 2001, peak value reduces 21% within than 1997.Due to mesh
The preceding disease there is no method to cure, and also without effective vaccine, therefore control HIV/AIDS prevalences of adopting an effective measure are current countries in the world
The highly important task faced.
The important component of AIDS prevention and control work is to find the infection sources.Exploitation effectively detection inhibition of HIV infection
Kit to AIDS prevention and control work be of great significance, HIV viral infections source can be cut off, be control HIV spread
Important means.
After carrying HIV, body can be directed to inhibition of HIV and produce corresponding antibody, therefore, the detection of HIV antibody
It is the important way for detecting inhibition of HIV infection.At present, detecting the major way of HIV antibody has ELISA, the experiment of glue particle agglutination
(PA), immunofluorescence experiment (IFA), immunoblotting analysis experiment (WB), radioimmunoprecipitation experiment (RIP), spot immune colloidal gold
Experiment and Chemiluminescence immunoassay etc..But there is for example complicated, costly, setting dependent on costliness for the above method
Standby, harsh experiment condition and the defects of professional testing staff.
Immuno-chromatographic test paper strip is a kind of rapid in-vitro diagnostic method that newly-developed gets up, and has had researcher at present
It is applied to the detection of HIV antibody.Compared with other detection methods, immuno-chromatographic test paper strip has simple, convenient fast
It is prompt, without instrument and equipment, it is cost-effective the advantages that.But existing immuno-chromatographic test paper strip is still suffered from when detecting HIV antibody
Certain defect, limits widely using for immuno-chromatographic test paper strip, including：(1) detection sensitivity is not high, weakly positive testing result
, easily there is false negative in unobvious；(2) current HIV antibody Test paper can only detect blood sample mostly, it is difficult to realize
The accurate detection of the viscous liquid sample such as mucous membrane of mouth diffusate.
In view of this, it is special to propose the present invention.
The content of the invention
The first object of the present invention is to provide a kind of test strips for detecting HIV antibody, and the test strips can amplify
The weakly positive testing result of HIV antibody improves the accuracy of detection so as to reduce false negative.
The second object of the present invention is to provide a kind of paper box of HIV antibody, and the kit not only includes foregoing inspection
Survey HIV antibody test strips, further include other detection components, can it is convenient, quickly and accurately detect HIV antibody.
In order to realize the above-mentioned purpose of the present invention, spy uses following technical scheme：
A kind of test strips for detecting HIV antibody, the test strips include bottom plate and are pasted onto on the bottom plate and take successively
Sample pad, label bonding pad, nitrocellulose filter and the water absorption pad connect；
Wherein, the sample pad is handled using sample pad treatment fluid, and the sample pad treatment fluid includes：PH7.2~7.5
PBS buffer, mass fraction be 0.5~1% polyethylene glycol, mass fraction be 0.5~1% casein, volume fraction
The PVP K-30 that Tween-20 and mass fraction for 0.3~1% are 0.6~1%；
The anti-human IgG antibodies of labeled substance markers are coated with the label bonding pad；
Detection line and nature controlling line are provided with the nitrocellulose filter, wherein, it is coated with HIV in the detection line and resists
Original, is coated with antiantibody on the nature controlling line, the antiantibody is with reference to the anti-human igg antibody.
The present invention relates to a kind of test strips for detecting HIV antibody, the sample pad of the test strips passes through special treatment fluid
Processing, the treatment fluid include PBS, polyethylene glycol, casein, Tween-20 and PVP K-30.Said components
Compound system hydrophilic, suspended dispersed and with strong buffer capacity is ultimately formed by cooperation each other.Use
Above-mentioned treatment fluid handles sample pad, by increasing capacitance it is possible to increase the hydrophily of sample pad, contributes to the quick humidification of sample pad, promotes chromatography
Effect occurs；Simultaneously as suspended dispersed effect and stablize pH environment, the sample pad can also in sample antibody
Play a protective role, and reduce non-specific binding.
In some specific embodiments, the label bonding pad includes label bonding pad 1 and label combines
Pad 2, the sample pad are overlapped on label bonding pad 1, and the label bonding pad 1 is overlapped on label bonding pad 2,
The label bonding pad 2 is overlapped on nitrocellulose filter, equal on the label bonding pad 1 and label bonding pad 2
The anti-human IgG antibodies of labeled substance markers are coated with evenly.
Label bonding pad in the above-mentioned test strips of the present invention includes the label bonding pads of two pieces of overlap joints, above-mentioned two pieces
Anti-human IgG antibodies have been evenly coated with label bonding pad.On the one hand, due to the increase of bonding pad, it is carried anti-human
The amount increase of IgG antibody, can strengthen it and capture the ability of antibody in sample, on the other hand, be combined with extending single labelled thing
The length of pad is compared so as to increase the mode of anti-human igg load capacity, and label bonding pad of the present invention mutually overlaps, and reduces
The total length of label bonding pad, it is long so as to causing test strips inconvenient to carry while cost to avoid the occurrence of test strips
The defects of rising.
In some specific embodiments, the label is selected from colloidal gold, electroselenium, latex particle, upper converting phosphor
Particle and fluorescent particle, it is preferable that the label is colloidal gold.
In some specific embodiments, the HIV antigens are selected from gp41, gp36 or combination.
In some specific embodiments, the coating concentration of the gp41 is 2.0~4.0mg/ml, such as 2.0,
2.5th, 3.0,3.5 or 4.0mg/ml.
In some specific embodiments, the coating concentration of the gp36 is 0.4~0.6mg/ml, such as 0.4,
0.45th, 0.5,0.55 or 0.6mg/ml.
In some specific embodiments, the coating concentration of the antiantibody is 0.3~0.5mg/ml, for example, 0.3,
0.35th, 0.4,0.45 or 0.5mg/ml.
In some specific embodiments, anti-human IgG antibodies dosage on the label bonding pad is 0.5
~0.9 μ g/cm2, for example, 0.5,0.6,0.7,0.8 or 0.9 μ g/cm2。
The present invention further limits the HIV antigens and is selected from gp41, gp36 or combination, therefore, of the present invention
Test strips can individually detect the antibody of AntiHIV1 RT activity 1 or the antibody of AntiHIV1 RT activity 2, alternatively, with the presence or absence of AntiHIV1 RT activity 1 in detection sample
At least one of antibody of antibody and AntiHIV1 RT activity 2.The present invention has further defined HIV antigens, antiantibody and anti-human igg and has resisted
The dosage of body, by the cooperation between above-mentioned three, further enhances the accuracy of test paper of the present invention.
In some specific embodiments, the anti-human IgG antibodies are mouse anti-human IgG antibodies, and the antiantibody is sheep
Dynamics.
In some specific embodiments, the test strips further include shell, and the shell has upper cover and lower cover to coordinate
Composition.
In some specific embodiments, the upper lid is equipped with thieff hatch and observation window, and the thieff hatch corresponds to sample
Product pad, the observation window correspond to nitrocellulose filter.
In some specific embodiments, the sample pad is selected from glass fibre element film, polyester film or filter paper fibre；
The label bonding pad is selected from glass fibre element film, polyester film or filter paper fibre；The water absorption pad is blotting paper.
In some specific embodiments, the polyethylene glycol in the sample pad treatment fluid is PEG 20000.
The invention further relates to a kind of kit for detecting HIV antibody, the kit includes any one of claim 1~8
The test strips and other detection components.Component needed for test strips and other detections is assembled into by kit of the present invention
Together, the convenience that immuno-chromatographic test paper strip uses can be dramatically increased.
In some specific embodiments, other described detection components include sample diluting liquid.
In some specific embodiments, other described detection components include sample diluting liquid, it is preferable that the sample
Product dilution includes the PBS buffer that pH is 7.2~7.5, the casein that mass fraction is 0.5~1.2%, mass fraction
The Tween-20 and volume fraction that 0.06~0.1% polyethylene glycol, volume fraction are 0.01~0.04% be 0.05~
0.15% Proclin-300.
The exposure of gum diffusate can solidify in atmosphere, cause sampling not carry out, so that cause detection to fail, therefore should
Gum diffusate is collected by this with dilution as early as possible.When mucous membrane of mouth diffusate sample by the method for immunochromatography into
During row detection, high viscosity substance present in sample can block the hole of perforated membrane.In addition, these high viscosity substances are also possible to lead to
Cross together with the epithelial cell combination to come off with human body, further block perforated membrane, cause immunochromatography to detect not smart
Really it can not even carry out.
Kit of the present invention includes special sample diluting liquid, the sample diluting liquid contain phosphate buffer,
The materials such as surfactant, casein and preservative.Wherein, phosphate buffer is biological cushion, casein can and sample
Non-specific material in this is combined together, that is, closes non-specific sites, can be to avoid sample and non-specific material
With reference to playing the role of physiology and mechanical protection, offset the false positive phenomenon as caused by non-specific material.Salt ion material is
Sodium chloride, salt ion can remove the stickum in sample, settle stickum, not influence sample on reagent strip
Flowing and infiltration.Surfactant can significantly change the interfacial tension between surface tension of liquid or two-phase.It is of the present invention dilute
Release liquid and also contain preservative, effectively extend the holding time of Sample dilution.Dilution of the present invention can also make sample exist
Fully pre-processed before detection, exclude to influence the unfavorable factor of detection reaction, improve the accuracy of detection.
In some specific embodiments, the polyethylene glycol in the sample diluting liquid is PEG 20000.
Compared with prior art, beneficial effects of the present invention are：
(1) sample pad of the present invention is handled through special treatment fluid, can strengthen the hydrophily of sample pad and to sample
The protective effect of middle antibody, while non-specific binding is reduced, therefore, compared with general HIV antibody Test paper, this hair
The bright test strips strengthen the sensitivity of detection, can reduce false negative when detecting weakly positive sample；
(2) label bonding pad of the present invention is 2 layers, and the anti-human IgG antibodies' that can strengthen labeled substance markers is negative
Carrying capacity, strengthens the sensitivity of detection, reduces false negative；
(3) kit of the present invention further includes sample buffer, the sample buffer contain phosphate buffer,
The stickum such as surfactant, casein and preservative, oral cavity diffusate through sample buffer of the present invention dilution after,
It can be used in the detection of HIV antibody test paper.
Brief description of the drawings
, below will be to tool in order to illustrate more clearly of the specific embodiment of the invention or technical solution of the prior art
Body embodiment or attached drawing needed to be used in the description of the prior art are briefly described, it should be apparent that, in describing below
Attached drawing be some embodiments of the present invention, for those of ordinary skill in the art, what is do not made the creative labor
Under the premise of, other attached drawings can also be obtained according to these attached drawings.
Fig. 1 is that the structure of the test strips of the mucous membrane of mouth diffusate HIV1/2 antibody fast detecting kits of the present invention is shown
It is intended to；
Fig. 2 is the sun of the mucous membrane of mouth diffusate HIV1/2 antibody fast detecting kits detection HIV antibody of the present invention
Property result, negative findings and null result schematic diagram；
Fig. 3 is kit described in kit described in embodiment 1 and comparative example 1 when detecting HIV1/2 negative antibody samples
Testing result, wherein above a row be test strips described in comparative example 1, below a row be test strips described in embodiment 1, it is each
What is added in 2 test strips of row is with a sample；
Fig. 4 is kit described in kit described in embodiment 1 and comparative example 1 when detecting HIV1/2 antibody positive samples
Testing result, wherein the test strips of a row (that is, drawing "×") are test strips described in comparative example 1 above, below a row (that is, draw
" √ ") be test strips described in embodiment 1, in 2 test strips of each row plus be with a sample, wherein 1#, 2#, 3# and
6# samples are weakly positive sample, and 2#, 4#, 5# and 7# sample are strong positive sample；
Fig. 5-A are detection when kit described in embodiment 1 detects 1# weakly positive samples with kit described in comparative example 1
As a result, wherein, the former (drawing "×") is test strips described in comparative example 1, and the latter's (drawing " √ ") is test strips described in embodiment 1；
Fig. 5-B are detection when kit described in embodiment 1 detects 2# weakly positive samples with kit described in comparative example 1
As a result, wherein, the former (drawing "×") is test strips described in comparative example 1, and the latter's (drawing " √ ") is test strips described in embodiment 1；
Fig. 5-C are detection when kit described in embodiment 1 detects 3# weakly positive samples with kit described in comparative example 1
As a result, wherein, the former (drawing "×") is test strips described in comparative example 1, and the latter's (drawing " √ ") is test strips described in embodiment 1；
Fig. 5-D are detection when kit described in embodiment 1 detects 6# weakly positive samples with kit described in comparative example 1
As a result, wherein, the former (drawing "×") is test strips described in comparative example 1, and the latter's (drawing " √ ") is test strips described in embodiment 1；
Fig. 6 is kit described in kit described in embodiment 1 and comparative example 1 when detecting HIV1/2 antibody positive samples
Testing result, wherein above a row (that is, drawing " √ ") be test strips described in embodiment 1, below a row (that is, drawing "×") be
Test strips described in comparative example 1, in 2 test strips of each row plus be with a sample, wherein sample #8, #11~#15, #
17 be strong positive sample；
Fig. 7 is kit described in kit described in embodiment 1 and comparative example 2 when detecting HIV1/2 negative antibody samples
Testing result, wherein above a row be test strips described in comparative example 2, below a row be test strips described in embodiment 1, it is each
What is added in 2 test strips of row is with a sample；
Fig. 8 is kit described in kit described in embodiment 1 and comparative example 2 when detecting HIV1/2 antibody positive samples
Testing result, wherein above a row be test strips described in comparative example 2, below a row be test strips described in embodiment 1, it is each
What is added in 2 test strips of row is that wherein 1#, 2#, 3# and 6# is weakly positive sample, and 4# and 5# are strong positive with a sample
Property sample；
Fig. 9-A are detection when kit described in embodiment 1 detects 1# weakly positive samples with kit described in comparative example 2
As a result, wherein, the former is test strips described in comparative example 2, the latter is test strips described in embodiment 1；
Fig. 9-B are detection when kit described in embodiment 1 detects 2# weakly positive samples with kit described in comparative example 2
As a result, wherein, the former is test strips described in comparative example 2, the latter is test strips described in embodiment 1；
Fig. 9-C are detection when kit described in embodiment 1 detects 3# weakly positive samples with kit described in comparative example 2
As a result, wherein, the former is test strips described in comparative example 2, the latter is test strips described in embodiment 1；
Fig. 9-D are detection when kit described in embodiment 1 detects 6# weakly positive samples with kit described in comparative example 2
As a result, wherein, the former is test strips described in comparative example 2, the latter is test strips described in embodiment 1；
Figure 10 is kit described in kit described in embodiment 1 and comparative example 2 when detecting HIV1/2 antibody positive samples
Testing result, wherein above a row be test strips described in comparative example 2, below a row be test strips described in embodiment 1, it is each
What is added in 2 test strips of row is same a sample, wherein, 7~8#, 11~15# and 17# samples are strong positive sample.
Embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
It will be appreciated that the following example is merely to illustrate the present invention, and it is not construed as limiting the scope of the invention.It is not specified in embodiment
Actual conditions person, the condition suggested according to normal condition or manufacturer carry out.Production firm is not specified in agents useful for same or instrument
Person, is that can buy the conventional products obtained by city.
Embodiment 1
A kind of method for the test strips and corresponding reagent box for preparing detection HIV1/2 antibody, including：
(1) 1L sample pad treatment fluids are prepared by formula as below：0.01M PBS (pH7.4), 6g PEG 20000s, 5g
Casein, 3mL Tween-20s, 6g PVP K-30s；
(2) 1L sample diluting liquids are prepared by formula as below：Disodium hydrogen phosphate dodecahydrate 2.9g, two hypophosphite monohydrate dihydros
Sodium 0.3g, sodium chloride 1.7g, casein 10g, PEG 20000 1g, Tween-20 0.2mL, Proclin-300 1ml,
Surplus is purified water；
(3) using aforementioned sample pad treatment fluid processing sample pad：The sample pad of every 31cm × 1.9cm, with 3.2ml institutes
State treatment fluid even spread to be placed on more than when drying 8 is small in electric drying oven with forced convection, kept dry is spare；
(4) by colloid gold label mouse anti-human IgG antibodies on glass fibre element film, coated weight is 0.7 μ g/cm2, drying；
(5) HIV antigens and sheep anti-mouse igg antibody are coated on nitrocellulose filter respectively, form detection line and Quality Control
Line, wherein, the concentration of gp41 is 3.0mg/ml in the HIV antigens, and the concentration of gp36 is 0.5mg/ml；The sheep anti-mouse igg
Antibody concentration is 0.4mg/ml；
(6) test strips are assembled in the following manner：Glass fibre element film, the nitric acid of sample pad, the label containing colloid gold particle
Cellulose membrane and blotting paper overlap successively, are pasted onto on bottom plate, and the sample pad is glass fibre membrane, the cellulose nitrate
The detection line of HIV recombinant antigens is coated with plain film and is coated with the nature controlling line of sheep anti-mouse igg antibody, the glass fibers
Colloid gold particle label on the plain film of dimension is colloid gold label mouse anti-human IgG antibodies, and the glass fibre element film is bilayer；
(7) test strips and prepared sample diluting liquid are assembled into kit.
Embodiment 2
The method that the test strips and corresponding reagent box of detection HIV1/2 antibody are prepared with reference to 1 the method for embodiment, difference
It is only that：
Prepare 1L sample pad treatment fluids formula be：0.01M PBS (pH7.4), 5g PEG 20000s, 10g junket eggs
In vain, 10mL Tween-20s, 10g PVP K-30s；
Prepare 1L sample diluting liquids formula be：Disodium hydrogen phosphate dodecahydrate 2.9g, two hypophosphite monohydrate sodium dihydrogens
0.3g, sodium chloride 1.7g, casein 5g, PEG 20000 0.6g, Tween-20 0.1mL, Proclin-300 0.5ml,
Surplus is purified water；
The coated weight of the colloid gold label mouse anti-human IgG antibodies is 0.5 μ g/cm2；The concentration of the gp41 is
4.0mg/ml；It is described；The concentration of gp36 is 0.4mg/ml；The sheep anti-mouse igg antibody concentration is 0.3mg/ml.
Embodiment 3
The method that the test strips and corresponding reagent box of detection HIV1/2 antibody are prepared with reference to 1 the method for embodiment, difference
It is only that,
Prepare 1L sample pad treatment fluids formula be：0.01M PBS (pH7.4), 10g PEG 20000s, 7g junket eggs
In vain, 7mL Tween-20s, 8g PVP K-30s；
Prepare 1L sample diluting liquids formula be：Disodium hydrogen phosphate dodecahydrate 2.9g, two hypophosphite monohydrate sodium dihydrogens
0.3g, sodium chloride 1.7g, casein 12g, PEG 20000 0.8g, Tween-20 0.4mL, Proclin-300
1.5ml, surplus are purified water；
The coated weight of the colloid gold label mouse anti-human IgG antibodies is 0.9 μ g/cm2；The concentration of the gp41 is
2.0mg/ml；It is described；The concentration of gp36 is 0.6mg/ml；The sheep anti-mouse igg antibody concentration is 0.5mg/ml.
Embodiment 4
The method that HIV1/2 antibody in sample is detected using kit of the present invention, is comprised the following steps：
(1) collecting sample, adds in 1ml dilutions and just can be used for detecting.Under room temperature, sample should after acquisition as early as possible into
Sample after processing, as can not be detected in time after sample process, should be placed in 2~8 DEG C of refrigerations, and sample should by row processing
It is completed after treatment in 10 days；If can not be detected in 10 days, Ying Yu -20 DEG C of preservations；(2) before detection by kit and
Sample is balanced to room temperature；(3) aluminum foil sack is torn, test is taken out and is placed on clean flat table top, aluminium foil bag is once torn
Open, test card must use immediately.Drawn with disposable plastic dropper and vertically dripped through the diluted sample solution of Sample dilution
Add 5~6 drops (about 100 μ L) in sample aperture.(4) observation testing result in 25-30 minutes.
Positive findings：It can be seen that two red stripes occur, one is located at detection line 5, and another at nature controlling line 6.
As long as any visible red stripes occur in detection line, no matter the depth is construed to positive.
Negative findings：Occur a red stripes, the redfree band at detection line 5 only at nature controlling line 6.
Null result：Do not occur red stripes at nature controlling line 6.
If testing result is the positive, other methods retest should be used again, and retest result is still positive sample
This, should send the HIV confirmatory tests room that national specific office determines to carry out confirmatory test.
Comparative example 1
Test strips and kit are prepared with reference to 1 the method for embodiment, are differed only in, the sample pad is without sample
The processing of present treatment liquid.
Comparative example 2
Test strips and kit are prepared with reference to 1 the method for embodiment, are differed only in, the mark containing colloid gold particle
The glass fibre membrane for remembering thing is individual layer.
Experimental example 1
The human immunity of National Institute for Food and Drugs Control's preparation is detected using kit described in the embodiment of the present invention 1
Defective virus (HIV) antibody mucous membrane of mouth diffusate National reference, specific detection method is with reference to embodiment 4, specific detection knot
Fruit is referring to table 1.According to testing result shown in table 1, kit of the present invention is in mucous membrane of mouth diffusate is detected
Meet national requirements during HIV1/2 antibody.
The result of HIV1/2 antibody in 1 kit of table detection mucous membrane of mouth diffusate
Experimental example 2
Using kit described in kit described in the embodiment of the present invention 1 and comparative example 1 respectively to negative sample, positive sample
This and weakly positive sample are detected, and specific detection method is referring to embodiment 4, and specific testing result is referring to Fig. 3~6.According to figure
Testing result shown in 3~6 understands that compared with the test paper handled without sample pad treatment fluid, the two is being examined test paper of the present invention
Indifference when surveying positive sample and negative sample, but when detecting weakly positive sample, test paper detection line colour developing of the present invention is bright
It is aobvious stronger than the detection line colour developing of test paper described in comparative example 1.
Experimental example 3
Using kit described in kit described in the embodiment of the present invention 1 and comparative example 2 respectively to negative sample, positive sample
This and weakly positive sample are detected, specific detection method referring to embodiment 4, specific testing result referring to Fig. 7~10, according to
Testing result shown in Fig. 7~10 understands that test paper of the present invention is compared with individual layer gold-labelled pad test paper, the two is in the positive sample of detection
Indifference when this and negative sample, but when detect weakly positive sample, test paper detection line colour developing of the present invention is substantially than contrasting
The detection line colour developing of test paper described in example 1 is strong.
Finally it should be noted that：The above embodiments are only used to illustrate the technical solution of the present invention., rather than its limitations；
Although the present invention is described in detail with reference to foregoing embodiments, it will be understood by those of ordinary skill in the art that：
It can still modify the technical solution described in foregoing embodiments, either to which part or whole technologies
Feature carries out equivalent substitution；And these modifications or replacement, the essence of appropriate technical solution is departed from each implementation of the present invention
The scope of example technical solution.
Claims (10)
1. a kind of test strips for detecting HIV antibody, it is characterised in that the test strips include bottom plate and are pasted onto on the bottom plate
And sample pad, label bonding pad, nitrocellulose filter and the water absorption pad overlapped successively；
Wherein, the sample pad is handled using sample pad treatment fluid, and the sample pad treatment fluid includes：The PBS of pH7.2~7.5
Casein that polyethylene glycol that buffer solution, mass fraction are 0.5~1%, mass fraction are 0.5~1%, volume fraction 0.3
~1% Tween-20 and mass fraction is 0.6~1% PVP K-30；
The anti-human IgG antibodies of labeled substance markers are coated with the label bonding pad；
Detection line and nature controlling line are provided with the nitrocellulose filter, wherein, HIV antigens, institute are coated with the detection line
State and antiantibody is coated with nature controlling line, the antiantibody is with reference to the anti-human IgG antibodies.
2. test strips according to claim 1, it is characterised in that the label bonding pad includes label bonding pad 1
With label bonding pad 2, the sample pad is overlapped on label bonding pad 1, and the label bonding pad 1 is overlapped on label
On bonding pad 2, the label bonding pad 2 is overlapped on nitrocellulose filter, and the label bonding pad 1 and label combine
The anti-human IgG antibodies of labeled substance markers are evenly coated with pad 2.
3. test strips according to claim 1, it is characterised in that the label is selected from colloidal gold, electroselenium, latex
Grain, upper converting phosphor particle and fluorescent particle, it is preferable that the label is colloidal gold.
4. test strips according to claim 1, it is characterised in that the HIV antigens are selected from gp41, gp36 or the group of the two
Close；Preferably, the coating concentration of the gp41 is 2.0~4.0mg/ml, and the coating concentration of the gp36 is 0.4~0.6mg/
ml。
5. test strips according to claim 1, it is characterised in that the coating concentration of the antiantibody is 0.3~0.5mg/
ml。
6. test strips according to claim 1, it is characterised in that the anti-human IgG antibodies are in the label bonding pad
Upper dosage is 0.5~0.9 μ g/cm2。
7. test strips according to claim 1, it is characterised in that the anti-human IgG antibodies are mouse anti-human IgG antibodies, institute
It is sheep anti-mouse igg antibody to state antiantibody.
8. test strips according to claim 1, it is characterised in that the test strips further include shell, on the shell has
Lid and lower cover, which are matched somebody with somebody, to be combined into；Preferably, the upper lid is equipped with thieff hatch and observation window, the thieff hatch counter sample pad, institute
State observation window and correspond to nitrocellulose filter.
9. a kind of kit for detecting HIV antibody, it is characterised in that the kit is included described in any one of claim 1~8
Test strips and other detection components.
10. kit according to claim 9, it is characterised in that other described detection components include sample diluting liquid, excellent
Selection of land, the sample diluting liquid include pH be 7.2~7.5 PBS buffer, mass fraction be 0.5~1.2% casein,
The Tween-20 and volume fraction that polyethylene glycol that mass fraction is 0.06~0.1%, volume fraction are 0.01~0.04% be
0.05~0.15% Proclin-300.
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| CN201711190830.5A CN107966561A (en) | 2017-11-24 | 2017-11-24 | A kind of test strips and kit for detecting HIV antibody |
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| CN201711190830.5A CN107966561A (en) | 2017-11-24 | 2017-11-24 | A kind of test strips and kit for detecting HIV antibody |
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| CN107966561A true CN107966561A (en) | 2018-04-27 |
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