CN207396503U - Detect the test strips and liquid detection cup of HIV antibody in urine - Google Patents
Detect the test strips and liquid detection cup of HIV antibody in urine Download PDFInfo
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- CN207396503U CN207396503U CN201721282596.4U CN201721282596U CN207396503U CN 207396503 U CN207396503 U CN 207396503U CN 201721282596 U CN201721282596 U CN 201721282596U CN 207396503 U CN207396503 U CN 207396503U
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Abstract
The utility model discloses a kind of test strips and liquid detection cup for detecting HIV antibody in urine, the test strips include bottom plate, and it is arranged at the sample pad on bottom plate, nitrocellulose filter and blotting paper, the both ends of the nitrocellulose filter overlap the sample pad and blotting paper respectively, the nearly nitrocellulose filter end is coated with the antibody of mouse anti-human igg of colloid gold label and the gp160 antigens of colloid gold label in the sample pad, the nitrocellulose filter is equipped with the first detection line of coating HIV recombinant antigen gp41 and HIV recombinant antigens gp160, and the control line of coating sheep anti-mouse antibody；For the test strips of the utility model by adding neoantigen gp160, the detection method than current anti-gp41 can reduce the generation of detection leakage phenomenon；The urine examination of the test strips high sensitivity of HIV antibody ensure that the real reliability of data in the detection urine of the utility model, available for the large-scale primary dcreening operation of AIDS.
Description
Technical field
The utility model belongs to pharmaceutical technology field, to resist more specifically, the utility model is related to HIV in a kind of detection urine
The test strips and liquid detection cup of body.
Background technology
Human immunodeficiency virus (Human Immunodeficiency Virus, HIV) is a kind of infection human immunity
The virus of system cells by destroying human immunocyte, causes immune system to lose resistance, and causes various diseases.State
Disease Control and Prevention Center of family statistics, from 2007, AIDS caused the highest infectious disease of death toll as China and continues into the present.Value
It is noted that China still has 32.1% the infected undiscovered at present.Determine a people whether aids infection poison it is unique
Method is to carry out AIDS virus detection.Data show, China's HIV detection limit rapid increases between 2008-2015.According to statistics, only
2015, the whole nation about had more than 1.43 hundred million people and receives HIV detections, accounts for the 10% of total population.
So large-scale HIV detections are all a stern challenge to human and material resources, financial resources, time, in particular with
" precisely, the global spread of AIDS, the infected risk example of medical staff propose the exploitation and innovation of HIV detection techniques
Quickly, it is noninvasive " requirement.
It is that primary dcreening operation is made a definite diagnosis to carry out westernblot again after the positive to be related to AIDS diagnostic program at present.Primary dcreening operation it is main
There are two types of detection methods：
1st, blood testing：Main method has immunochromatographic method (colloidal gold method or emulsion method), chemoluminescence method, time resolution
Fluorescence method gathers blood sample of patient, for the human immune deficiency in external qualitative or quantitative detection human serum, blood plasma and whole blood
Viral (HIV-1/HIV-2) antibody.These methods are mainly that the presence or absence of HIV antibody by detecting suspicious person blood judges whether
Infection.
But the method for blood testing is a kind of traumatic blood testing, and blood born is also 3 big ways of AIDS propagation
One of footpath, wherein, chemiluminescence and time-resolved fluorescence need large scale equipment, and hospital only at county level and above is equipped with.Secondly,
Time-consuming for this 2 kinds of methods, it is necessary to special operating personnel and training.The common drawback of three is：
A, the risk that medical staff is stabbed and infected during blood drawing；
B, various medical instrument rubbish and processing after drawing blood are (such as：Syringe needle, acupuncture needle, dental appliance, beauty treatment instrument etc.)
There are the risks of infection；
C, the blood sample of acquisition needs to centrifuge, it is difficult to carry out self detection of patient so as to ensure the privacy of patient.
2nd, oralmucosal transudate fluid detects：Main method has immunochromatographic method (colloidal gold method or emulsion method).Use oral cavity
Diffusate of the swab after gum line constantly wipes detects the HIV-1/2 type human immune deficiencies in human oral mucosa's diffusate
Antiviral antibody.
The method is a kind of minimally invasive detection, much smaller compared to the risk of blood born, but there are still the risk such as AIDS of propagation
The people at highest risk of exposed population group.
Antibody in urine there are HIV specificity is by numerous studies and confirmation.Nothing/extremely weak communicable urine detection
Method CFDA approvals only have enzyme-linked immunization at present, and time-consuming (3h), and can only qualitative detection.Qualitative detection can equally be carried out
Colloidal gold method since its is easy to operate, it is short and deposited by pro-gaze and the first choice of consideration, but since urine antibody content is very low to take
The sensitivity the problem of, therefore existing market does not find similar product also, in addition, having been reported that, urine detection is compared to blood, saliva
In the presence of higher false positive issue.
Utility model content
Based on this, the defects of in order to overcome the above-mentioned prior art, the utility model provides HIV in a kind of detection urine and resists
The test strips and detection cup of body.
In order to realize above-mentioned purpose of utility model, the utility model takes following technical scheme：
The test strips of HIV antibody in a kind of detection urine, the test strips include bottom plate and the sample being arranged on bottom plate
Product pad, nitrocellulose filter and blotting paper, the both ends of the nitrocellulose filter overlap the sample pad and blotting paper, institute respectively
State antibody and colloid gold label that the nearly nitrocellulose filter end in sample pad is coated with the mouse anti-human igg of colloid gold label
Gp160 antigens and formed gold mark layer, the nitrocellulose filter be equipped with coating HIV recombinant antigen gp41 and HIV recombinant antigens
The first detection line of gp160 and the control line for being coated with sheep anti-mouse antibody.
In wherein some embodiments, the second of coating HIV recombinant antigens gp36 is additionally provided on the nitrocellulose filter
Detection line, the nearly nitrocellulose filter end is also coated with the gp36 antigens of colloid gold label in the sample pad.
In wherein some embodiments, one end of the gold mark closely described nitrocellulose filter of layer and first detection line
The distance between for 4~7.5mm, be preferably 6.5mm.
In wherein some embodiments, the one end of the sample pad away from the nitrocellulose filter with it is described gold mark layer it
Between distance for 20~25mm, be preferably 22mm.
In wherein some embodiments, the distance between first detection line and second detection line are 3~4mm,
The distance between second detection line and the control line be 3~4mm, the closely described nitrocellulose filter end of the blotting paper with
The distance between described second detection line is 6~7mm, and the length of the blotting paper is 25~30mm.
In wherein some embodiments, the distance between first detection line and second detection line be 3.5mm, institute
The distance between the second detection line and the control line are stated as 3.5mm, one end of the closely described nitrocellulose filter of the blotting paper
The distance between described second detection line is 6.5mm, and the length of the blotting paper is 28mm.
In wherein some embodiments, the concentration of the colloid gold label gp160 antigens is 1.5~3.5 μ g/ml；It is described
The concentration of the antibody of colloid gold label mouse anti-human igg is 4.5~6.5 μ g/ml；The concentration of the colloid gold label gp36 antigens is
4.5~6.5 μ g/ml；The dosage of the colloid gold label raw material is 2~3.5ul/cm；The bag of the HIV recombinant antigens gp41
It is 0.5-2.0mg/ml by concentration, the coating concentration of the HIV recombinant antigens gp160 is 0.2-0.8mg/ml, the HIV restructuring
The coating concentration of antigen gp36 is 0.8-1.0mg/ml；The dosage of the HIV recombinant antigens is 0.12-0.15ul/mm, described
The concentration of sheep anti-mouse igg antibody is 0.8-1.0mg/ml, and the dosage of the sheep anti-mouse igg antibody is 0.09-0.22ul/mm.
In wherein some embodiments, the sample pad is handled using sample pad treatment fluid, the sample pad processing
Liquid includes buffer solution, aqueous auxiliary rheological agents and the sealer that pH is 8~11, wherein, compared with the buffer solution of every 100mL,
The additive amount of the aqueous auxiliary rheological agents is 0.05~10g, and the additive amount of the sealer is 0.05~10g.
In wherein some embodiments, the sample pad treatment fluid further includes the Na of 0.01~1M2CO3Or K2CO3。
In wherein some embodiments, S9 is further included in the sample pad treatment fluid；Compared with the buffering of every 100mL
Liquid, the additive amount of the S9 is 0.01~1g.
In wherein some embodiments, the aqueous auxiliary rheological agents for hydroxypropyl methyl cellulose, in PAA, PEO at least
It is a kind of.Preferably, the aqueous auxiliary rheological agents for hydroxypropyl methyl cellulose and PAA mixture, wherein, hydroxypropyl methyl is fine
Dimension element accounts for the 45~70% of the total weight of the mixture.
In wherein some embodiments, the sealer is rabbit anteserum.
In wherein some embodiments, it is described coating buffer solution include pH be 7-8,1-50mM Tris-HCl buffer solutions,
S9 and urea；Wherein, compared with the Tris-HCl buffer solutions of every 100mL, the additive amount of the S9 is 0.01~1g, and the urea adds
Dosage is 2~10g.
The utility model additionally provides a kind of test strips for detecting HIV antibody in urine, and the test strips include bottom plate, with
And be sequentially arranged in sample pad, gold-labelled pad, nitrocellulose filter and blotting paper on bottom plate, one end of the gold-labelled pad is overlaid on
Under the sample pad, the other end of the gold-labelled pad and one end of blotting paper are overlapped on the nitrocellulose filter respectively
On, the antibody of the mouse anti-human igg of colloid gold label and the gp160 antigens of colloid gold label are coated in the gold-labelled pad and is formed
Gold mark layer, the nitrocellulose filter are equipped with the first detection of coating HIV recombinant antigen gp41 and HIV recombinant antigens gp160
Line and the control line for being coated with sheep anti-mouse antibody.
In wherein some embodiments, the second of coating HIV recombinant antigens gp36 is additionally provided on the nitrocellulose filter
Detection line is also coated with the gp36 antigens of colloid gold label in the gold-labelled pad.
In wherein some embodiments, one end of the gold mark closely described nitrocellulose filter of layer and first detection line
The distance between for 4~7.5mm, be preferably 6.5mm.
In wherein some embodiments, the one end of the sample pad away from the nitrocellulose filter with it is described gold mark layer it
Between distance for 20~25mm, be preferably 22mm.
In wherein some embodiments, the distance between first detection line and second detection line are 3~4mm,
The distance between second detection line and the control line be 3~4mm, the one of the closely described nitrocellulose filter of the blotting paper
The distance between end and second detection line are 6~7mm, and the length of the blotting paper is 25~30mm.
In wherein some embodiments, the distance between first detection line and second detection line be 3.5mm, institute
The distance between the second detection line and the control line are stated as 3.5mm, the closely described nitrocellulose filter end of the blotting paper and institute
The distance between second detection line is stated as 6.5mm, the length of the blotting paper is 28mm.The utility model additionally provides a kind of liquid
Body detection cup, including cup body, test strip clamp plate and cover film, at least a portion of the test strip clamp plate is placed in cup body, is set thereon
There is test paper slot, the test strips of HIV antibody in above-described detection urine, the cover film are placed in the test paper slot
The test strips surface is covered in by test paper slot opening face.
In wherein some embodiments, space is left between the test strip clamp plate and the cup body, the test strip clamp plate is hung down
Directly extend outwardly in the bottom of cup and stretched out from the cup body, the top edge of the cup body is in the detection urine
6~15mm of gold mark layer of the test strips of HIV antibody.
In wherein some embodiments, the test paper slot is equipped with several, and Poison is also placed in the test paper slot
Test strip, the illicit drugs inspection test strips are positioned over different test paper from the test strips of HIV antibody in the detection urine
In slot.
In wherein some embodiments, length of the cover film bottom edge apart from test strips bottom margin is the examination
The 1/6-5/6 of the sample pad length of paper slip.
In wherein some embodiments, the cover film is hyaline membrane.
In order to solve the problems, such as the sensitivity and specificity of urine detection HIV, the utility model people of the utility model implements
A variety of researchs, through determining with solid-phase immunity chromatographic theory, using the sensibility processing of early period sample pad and specific antigen
HIV antibody in gp160, double-antibody method indirect method joint technology detection urine, can improve sensitivity and specificity.If urine
Contain HIV antibody in liquid sample, then HIV antibody is combined with colloid gold particle label, forms compound, and is diffused into nitric acid fibre
It is further chromatographed on the plain film of dimension, when running into the pairing antigen being coated at detection zone on nitrocellulose filter (T lines), compound
It then combines, is trapped at coating with envelope antigen again, when captured compound reaches certain amount, then forming naked eyes can
The T lines seen, illustrate to contain HIV antibody in sample, if not occurring, illustrate that sample is less than the minimum inspection of test strips for negative or content
Survey limit.Quality control standard of the control zone (C lines) as test strips, positive and negative sample are present with when detecting.
Compared with prior art, the utility model has the advantages that：
1st, by adding neoantigen gp160, the detection method than current anti-gp41 more carries the test strips of the utility model
Preceding precognition HIV infection, the periodic regime of detection is wider, as mark, can capture the antibody in addition to IgG types, conduct
Coating can make up the HIV antibody of gp41 missing inspections；
2nd, the test strips of HIV antibody reduce HIV contactees (such as medical work people in the detection urine of the utility model
Member, pre-marital or pregnant woman inspection etc.) due to the risk that collecting sample is stabbed and is infected, urine sample need not be concentrated and pre-processed,
Storage, convenient transportation, risk of leakage is low, without more processing equipment, without carrying out the technical ability training of specialty again to medical staff
Instruction；Meanwhile with the development of medical treatment, OTC detections are had increasingly been towards, release hospital, the pressure of medical staff significantly.Additionally
Personal formula detection can be carried out, it is easy to operate, individual privacy is protected, reaches real POCT detections；
3rd, the urine examination of the test strips high sensitivity of HIV antibody ensure that the true of data in the detection urine of the utility model
Reliability, available for the large-scale primary dcreening operation of AIDS.
Description of the drawings
Fig. 1 be the utility model detection urine in HIV antibody test strips an embodiment structure diagram；
Fig. 2 be the utility model detection urine in HIV antibody test strips another embodiment structure diagram；
Fig. 3 is the structure diagram of an embodiment of the liquid detection cup of the utility model；
Fig. 4 is the structure diagram of another embodiment of the liquid detection cup of the utility model；
Reference numeral：1st, sample pad；2nd, label pad；3rd, bottom plate；4th, nitrocellulose filter；5th, control line；61st, detection line
T1；62nd, detection line T2；7th, blotting paper；1', cup body；2', test strip clamp plate；21', test paper slot；3', cup lid；31', sealing ring.
Specific embodiment
The utility model is further discussed below in the following with reference to the drawings and specific embodiments, and the utility model is not addressed part and is applicable in
In the prior art.Specific embodiment of the utility model is given below, but embodiment is merely to be described in further detail this theory
It is bright, it is not intended to limit the claim of the utility model.Reagent or raw material used in following embodiment, unless otherwise specified,
It derives from commercially available.
Referring to Fig. 1, in the detection urine of an embodiment HIV antibody test strips, including bottom plate 3 and set
In sample pad 1, nitrocellulose filter 4 and blotting paper 7 on bottom plate, the both ends of the nitrocellulose filter 4 overlap described respectively
Sample pad 1 and blotting paper 7, the nearly nitrocellulose filter end is coated with the mouse anti-human igg of colloid gold label in the sample pad 1
Antibody and colloid gold label gp160 antigens and formed gold mark layer, the nitrocellulose filter 4 be equipped with coating HIV restructuring
The first detection line 61 of antigen gp41 and HIV recombinant antigen gp160 and the control line 5 for being coated with sheep anti-mouse antibody.
Specifically, in the present embodiment, it is additionally provided with coating HIV recombinant antigens gp36's on the nitrocellulose filter 4
Second detection line 62, the nearly nitrocellulose filter end is also coated with the gp36 antigens of colloid gold label in the sample pad 1.This
The setting of sample may be such that test strips detect the infection of HIV-1 and HIV-2 types simultaneously, due to two detection lines respectively for HIV-1 and
HIV-2 types infect, and can more intuitively find out HIV infection type.
Further, it is described gold mark the closely described nitrocellulose filter of layer one end and first detection line 61 between away from
From for 4~7.5mm, being preferably 6.5mm；One end and first detection line of the gold mark closely described nitrocellulose filter of layer it
Between distance can influence the detection accuracy of weakly positive sample, if apart from too small, be easier to weak positive sample occur to show false the moon
If the problem of effect, apart from excessive, can influence detection efficiency；When distance is 6.5mm, detection result is optimal.
Further, the distance between layer is marked in the one end of the sample pad 1 away from the nitrocellulose filter with the gold
It is preferably 22mm for 20~25mm；If the sample pad away from the nitrocellulose filter end and it is described gold mark layer between away from
From too small, since urine impact force is larger, the colloid gold label object in gold mark layer is easily broken up so that part colloid gold label object
It can be broken up to sample-adding side rather than be chromatographed onto nitrocellulose filter, so as to influence the stability of testing result；Apart from excessive
Loading efficiency can then be influenced.When distance is 22mm, detection result is optimal.
Further, the distance between first detection line 61 and second detection line 62 are 3~4mm, described the
The distance between two detection lines 62 and the control line 5 are 3~4mm, the nearly nitrocellulose filter end of the blotting paper 7 and institute
The distance between second detection line 62 is stated as 6~7mm, the length of the blotting paper 7 is 25~30mm.Specifically, described first
The distance between detection line 61 and second detection line 62 are 3.5mm, between second detection line 62 and the control line 5
Distance for 3.5mm, the distance between the nearly nitrocellulose filter end of the blotting paper 7 and described second detection line 62 are
6.5mm, the length of the blotting paper 7 is 28mm.
Referring to Fig. 2, in the detection urine of another embodiment HIV antibody test strips, and in the above embodiment
Detection urine in HIV antibody test strips the difference is that：It is additionally arranged gold-labelled pad 2, and gold mark layer is arranged on gold-labelled pad 2
On, one end of the gold-labelled pad 2 is overlaid under the sample pad 1, the other end of the gold-labelled pad 22 and the one of blotting paper 7
End is overlapped on respectively on the nitrocellulose filter 4.
Refer to Fig. 3~Fig. 4, the liquid detection cup of an embodiment, including cup body 1', test strip clamp plate 2' and cover film,
At least a portion of the test strip clamp plate 2' is placed in cup body 1', which is provided in test paper slot 21', the test paper slot 21'
The test strips of HIV antibody in above-described detection urine are placed with, the cover film is covered in by test paper slot 21' openings face
The test strips surface.
Specifically, in the present embodiment, the test paper slot 21' is equipped with several, is also put in the test paper slot 21'
Illicit drugs inspection test strips are equipped with, the test strips of the illicit drugs inspection test strips and HIV antibody in the detection urine are positioned over not
In same test paper slot 21'.
More specifically, length of the cover film bottom edge apart from test strips bottom margin is the sample of the test strips
Pad the 1/6-5/6 of length.
As shown in figure 3, in one embodiment, the test strip clamp plate 2' is all placed in cup body 1', the liquid detecting
Cup further includes cup lid 3', cup lid 3' and cup body 1' thread fittings, and cup lid 3' bottoms are equipped with sealing ring 31'.
As shown in figure 4, in one embodiment, the test strip clamp plate 2' is partially disposed in cup body 1', the test strip clamp plate
2' extends outwardly perpendicular to the cup body 1' bottoms and is stretched out from the cup body 1', and the part that test strip clamp plate 2' stretches out can facilitate hand
It holds；Space is left between the test strip clamp plate 2' and the cup body 1', in order to accept urine；The top edge of the cup body 1'
6~15mm of gold mark layer of the test strips of HIV antibody in the detection urine.
Illustrate the other improvements of the utility model part with reference to specific embodiment.
Embodiment 1
Referring to Fig. 1, in a kind of detection urine of the present embodiment HIV antibody test strips, the test strips include bottom plate
3 and the sample pad 1 and nitrocellulose filter 4 that are arranged on bottom plate 3, the sample pad 1 is using at sample pad coating buffer
Reason, the both ends of the nitrocellulose filter 4 are overlapped with sample pad 1 and blotting paper 7, colloid are coated in the sample pad 1 respectively
The antibody of gold mark mouse anti-human igg, colloid gold label gp160 antigens and colloid gold label gp36 antigens, the nitrocellulose
Film 4 is equipped with the detection line 61 of coating HIV recombinant antigen gp41 and HIV recombinant antigens gp160, coating HIV recombinant antigens gp36
Detection line 62 and be coated with sheep anti-mouse antibody control line 5.
Wherein, the sample pad treatment fluid include pH for 9, the Tris-HCl buffer solutions of 100mM, HPMC, PAA, rabbit anteserum,
S9 and 0.09M Na2CO3；Wherein, compared with the Tris-HCl buffer solutions of every 100mL, the additive amount of HPMC is 0.1g, and PAA's adds
Dosage is 0.1g, and the additive amount of rabbit anteserum is 0.1g, and the additive amount of S9 is 0.1g.
First detection line 61 is after diluting recombinant antigen to working concentration by coating buffer solution, to line cellulose nitrate
On film 4, which includes pH for 7.4,20mM Tris-HCl buffer solutions and S9 and urea；Wherein, compared with every
The Tris-HCl buffer solutions of 100mL, the additive amount of the S9 is 0.1g, and the additive amount of the urea is 5g.
The test strips can be used for detecting HIV-1 types and the infection of HIV-2 types.
In this embodiment, the concentration of the colloid gold label gp160 antigens is 2.5 μ g/ml；The colloid gold label mouse
The concentration of the antibody of anti-human igg is 5.6 μ g/ml；The concentration of the colloid gold label gp36 antigens is 5.6 μ g/ml.
In this embodiment, the dosage of the colloid gold label raw material is 2.75ul/cm；
In this embodiment, the coating concentration of the HIV recombinant antigens gp41 be 1.0mg/ml, the HIV recombinant antigens
The coating concentration of gp160 is 0.6mg/ml, and the coating concentration of the HIV recombinant antigens gp36 is 0.6mg/ml, the HIV restructuring
The dosage of antigen is 0.12ul/mm.
In this embodiment, the concentration of the sheep anti-mouse antibody is 1.0mg/ml, and the dosage of the sheep anti-mouse antibody is
0.10ul/mm。
The preparation method of the test strips of HIV antibody is as follows in the detection urine of the embodiment：
1) sample pad treatment fluid, is prepared, is coated on glass fibre element film, coating concentration 40ul/cm2, 25 DEG C are placed in,
Sample pad is made in humidity 10%~30%, drying and processing 18-22h；
2) mouse IgG and colloid gold particle, are combined to form into colloid gold label mouse IgG antibody probe；Gp160 antigens and colloid
Gold grain mark to be formed colloid gold label HIV specific antigens probe (before gp160 antigens are combined with colloid gold particle, it is necessary to
Gp160 antigens are dissolved in HIV coating dilutions, the HIV coatings dilution includes pH for the 7.4, Tris-HCl of 20mM
Buffer solution, S9 and urea；Wherein, compared with the Tris-HCl buffer solutions of every 100mL, the additive amount of the S9 is 0.1g, the urea
Additive amount be 5g)；Gp36 antigens mark to form colloid gold label HIV specific antigen probes with colloid gold particle.By above-mentioned 3
Kind gold grain mixing, with gold spraying instrument even application in sample pad, metal spraying width 6mm, length 2.8cm, 25 DEG C are dried 18-22h,
Humidity 10%~30%, for use；
3) HIV specific antigen gp41 and gp160, is diluted to working concentration with coating buffer solution, is carried out with Membrane jetter
It lines and the first detection line is formed on nitrocellulose filter, by the HIV specific antigens gp36 Tris- that pH is 7.4,20mM
HCl buffer solutions are diluted to working concentration, with Membrane jetter line the second detection line is formed on nitrocellulose filter, by goat-anti
Mouse antibody is diluted to working concentration, and formation C nature controlling lines line on nitrocellulose filter with Membrane jetter, in 25 DEG C, humidity
10~30%, drying and processing, 18-22h；
4), by sample pad, nitrocellulose filter and blotting paper be sequentially overlapped on bottom plate to get.
Embodiment 2
The test strips structure of HIV antibody and preparation method are same as Example 1 in the detection urine of the present embodiment, different
Be：The concentration of the colloid gold label gp160 antigens is 1.5 μ g/ml；The antibody of the colloid gold label mouse anti-human igg
Concentration is 4.5 μ g/ml；The concentration of the colloid gold label gp36 antigens is 4.5 μ g/ml.The use of the colloid gold label raw material
It measures as 2ul/cm；The coating concentration of the HIV recombinant antigens gp41 be 0.5mg/ml, the coating of the HIV recombinant antigens gp160
Concentration is 0.2mg/ml, and the coating concentration of the HIV recombinant antigens gp36 is 0.8mg/ml, the dosage of the HIV recombinant antigens
It is 0.15ul/mm.The concentration of the sheep anti-mouse antibody is 0.8mg/ml, and the dosage of the sheep anti-mouse antibody is 0.2ul/mm.
The sample pad treatment fluid of the present embodiment includes pH for 10, the Tris-HCl buffer solutions of 10mM, HPMC, PAA, rabbit blood
Clearly, S9 and 0.5MNa2CO3；Wherein, compared with the Tris-HCl buffer solutions of every 100mL, the additive amount of HPMC is 0.7g, PAA's
Additive amount is 0.3g, and the additive amount of rabbit anteserum is 1g, and the additive amount of S9 is 1g.
The coating buffer solution of the present embodiment includes pH for 7,10mM Tris-HCl buffer solutions and S9 and urea；Wherein, phase
For the Tris-HCl buffer solutions of every 100mL, the additive amount of the S9 is 0.01g, and the additive amount of the urea is 2g.
Embodiment 3
The test strips structure of HIV antibody and preparation method are same as Example 1 in the detection urine of the present embodiment, different
Be：The concentration of the colloid gold label gp160 antigens is 3.5 μ g/ml；The antibody of the colloid gold label mouse anti-human igg
Concentration is 6.5 μ g/ml；The concentration of the colloid gold label gp36 antigens is 6.5 μ g/ml.The use of the colloid gold label raw material
It measures as 3.5ul/cm；The coating concentration of the HIV recombinant antigens gp41 be 2.0mg/ml, the bag of the HIV recombinant antigens gp160
It is 0.8mg/ml by concentration, the coating concentration of the HIV recombinant antigens gp36 is 0.9mg/ml, the use of the HIV recombinant antigens
Amount is 0.13ul/mm.The concentration of the sheep anti-mouse antibody is 0.9mg/ml, and the dosage of the sheep anti-mouse antibody is 0.15ul/
mm。
The sample pad treatment fluid of the present embodiment includes pH for 11, Tris-HCl buffer solutions, HPMC, rabbit anteserum, the S9 of 200mM
With 1M Na2CO3；Wherein, compared with the Tris-HCl buffer solutions of every 100mL, the additive amount of HPMC is 5g, the additive amount of rabbit anteserum
For 1g, the additive amount of S9 is 0.5g.
The coating buffer solution of the present embodiment includes pH for 8,50mM Tris-HCl buffer solutions and S9 and urea；Wherein, phase
For the Tris-HCl buffer solutions of every 100mL, the additive amount of the S9 is 1g, and the additive amount of the urea is 10g.
Embodiment 4
Referring to Fig. 2, the test strips structure of the present embodiment is identical with the test strips structure of embodiment 1, unlike, it sets
There is label pad, and colloid gold particle is sprayed in label pad.
5 liquid detection cup of embodiment
Referring to Fig. 3, a kind of liquid detection cup of the present embodiment, including cup body 1', test strip clamp plate 2', cover film, cup lid
3' and the sealing ring 31' being arranged in cup lid 3', the test strip clamp plate 2' are placed in cup body 1', which is provided with test paper slot
The test strips of HIV antibody in the detection urine of embodiment 1 are placed in 21', the test paper slot 21', the cover film is by trying
Paper slot 21' openings face is covered in the test strips surface, length of the cover film bottom edge apart from test strips bottom margin
Spend the 1/6-5/6 of the sample pad length for the test strips.
Tests below example 1~4 is the formulation screening test example of the sample pad treatment fluid of the test strip of the present invention.
The Selection experiment of the auxiliary agent of 1 sample pad treatment fluid of test example
The one or more auxiliary rheological agents of addition can adjust the rheological characteristic of liquid, prevent gold grain from drying up, this test example root
Aqueous auxiliary rheological agents are selected according to urine feature, according to its cellulose family, polyoxyethylene, polyaerylic acid class, natural gum and its are changed
The classification of property object have chosen respectively 1., hydroxypropyl methyl cellulose 0.1%, 1%, 10%；2., PEO 0.1%, 1%, 10%；
3., PAA0.1%, 1%, 10%；4., sodium alginate 0.1%, 1%, 10% (wherein, hydroxypropyl methyl cellulose 0.1% refers to
The buffer solution 0.1g hydroxypropyl methyl celluloses of pH9.00.1M Tris-HCl per 100mL, in the identical item of solvent
Under part, remaining concentration and so on), it is respectively coated on glass fibre element film, coating concentration 40ul/cm2, it is placed in 25
DEG C, humidity 10%~30%, drying and processing, 18-22h, obtained sample pad.
The spraying of gold grain and the preparation method of nitrocellulose membrane are the same as embodiment 1.
The preparation method of test strips is the same as embodiment 1.
The test strips that this test example is prepared are tested as follows：
Selection Healthy People compares, the patient that makes a definite diagnosis of the detection HIV positives and with Healthy People feminine gender urine and the HIV positives
Strong sun, middle-jiao yang, function of the spleen and stomach, the internal reference of weak sun is made in Urine in Patients.2 drops (60-80ul) sample is added dropwise respectively to originally with rubber head dropper
In the sample pad of test strips made from test example, due to capillarity, sample will be fine along test strips to gold grain and nitric acid
The plain film movement of dimension, treats that sample is completely dissolved gold grain and to nitrocellulose membrane flow, as a result starts to show；It is observed after 15 minutes
Show result (note：It is invalid to develop the color after 30 minutes).Test result is as shown in table 1.
Influence of the different aqueous auxiliary rheological agents of table 1 to sample
Note：Totally 10 colour developing gradients：C1~C9, B.Wherein B represents " blank ", no band；C1~C9 represents that colour developing is strong
Degree, wherein number is bigger to represent that colour developing is more shallow.C8+ represents to develop the color between C8~C7, other and so on.
As can be known from Table 1, different types of auxiliary rheological agents effect is hydroxypropyl methyl cellulose successively at low concentrations
＞ PEO ≈ PAA ＞ sodium alginates.Compared with other aqueous auxiliary rheological agents, hydroxypropyl methyl cellulose more effectively weakens
The flowing of moisture, while give the liquid phase reactor environment of gold grain abundance so that reaction fully carries out, and improves sensitivity.
PEO and PAA has lockked the flowing of moisture to a certain extent, but the concentration ratio hydroxypropyl methyl cellulose used is higher.
Sodium alginate effect is most weak.Therefore it is preferred that hydroxypropyl methyl cellulose, alternative polyoxyethylene, polyaerylic acid class.In addition, inventor
It has been unexpectedly discovered that hydroxypropyl methyl cellulose and PAA are used as aqueous auxiliary rheological agents and in suitable concentration model when simultaneously
When enclosing (namely hydroxypropyl methyl cellulose accounts for the 80~90% of the two total weight, similarly hereinafter), the two plays synergistic effect,
Sensitivity is can further improve, and gold grain will not be caused to be separated from water.
The elimination experiment of 2 false positive of test example
Urine pH major part slant acidity, pH is very sensitive for the detection of urine, and gold grain is easily sent out in acid condition
Raw deposition, and sodium carbonate can finely tune the acidic materials (uric acid, creatine etc.) in urine.Sodium ion therein can also remain anti-
Antigen-antibody reaction is required ionic condition, ensure that the specific reaction of Ag-Ab.
By the Na of various concentration gradient2CO3(as shown in table 2) is incorporated in pH9.0,0.1M Tris-HCL buffer solutions, so
After be coated on glass fibre element film, coating concentration 40ul/cm2, it is placed in 25 DEG C, humidity 10%~30%, drying and processing 18-
Sample pad is made in 22h.
The spraying of gold grain and the preparation method of nitrocellulose membrane are the same as embodiment 1.
The preparation method of test strips is the same as embodiment 1.
The test strips that this test example is prepared are tested, and test method is the same as test example 1, test result such as 2 institute of table
Show.
2 Na of table2CO3Influence of the content to sample
Note：Totally 10 colour developing gradients：C1~C9, B.Wherein B represents " blank ", no band；C1~C9 represents that colour developing is strong
Degree, wherein number is bigger to represent that colour developing is more shallow.C8+ represents to develop the color between C8~C7, other and so on.
From table 2 it can be seen that part false positive can lead to existing for the alkaline buffer system of pH9.00.1M Tris-HCL
The specificity for adding in sodium carbonate guarantee reaction is crossed, while introduces certain Na ion concentration and maintains immunochromatography process moderate resistance
The ion that antigen-antibody reaction needs.
The Selection experiment of albumen in 3 sample pad treatment fluid of test example
Protein is practically free of in urine, add in a little closed protein not only can to avoid occur non-specific binding, and
And the environment in the stabilization urine reaction of urine molecule total amount can be increased.The closed protein rabbit anteserum can be with as routine BSA
It is combined with the albumen non-specific sites in sample, great advantage is can to close endogenic Fc segments in sample, resistance
Disconnected antibody is combined with the Fc receptors in sample, reduces background, reduces false positive.
Liquid based on preparation pH9.0,0.1M Tris-HCL buffer solutions, adds hydroxypropyl methyl cellulose 0.1%,
PAA0.1%, 0.09MNa2CO3, then it is coated on glass fibre element film, coating concentration 40ul/cm2, 25 DEG C are placed in, humidity
Sample pad is made in 10%~30%, drying and processing 18-22h.
The spraying of gold grain and the preparation method of nitrocellulose membrane are the same as embodiment 1.
The preparation method of test strips is the same as embodiment 1.
The test strips that this test example is prepared are tested, and test method is the same as test example 1, test result such as 3 institute of table
Show.
The comparison that 3 rabbit anteserum of table, BSA influence sample
Note：Totally 10 colour developing gradients：C1~C9, B.Wherein B represents " blank ", no band；C1~C9 represents that colour developing is strong
Degree, wherein number is bigger to represent that colour developing is more shallow.C8+ represents to develop the color between C8~C7, other and so on.In addition, in upper table
The numerical value representative before the % of rabbit anteserum dosage is characterized compared with the buffer solution of every 100mL, the quality dosage (g) of rabbit anteserum；On
Numerical value before the % of characterization BSA dosages in table represents the buffer solution compared with every 100mL, the quality dosage (g) of BSA.
From table 3 it can be seen that rabbit anteserum is better than BSA.BSA is widely used in blood, blood plasma as sealer as envelope
Albumen is closed, is also commonly used in Antibody preparation, and the antibody in gold grain is a small amount of due to that may contain among the process of expression
The antibody of anti-BSA do not remove, it is caused to carry out reaction with BSA and causes above-mentioned vacation sun.And there is no the anti-of anti-rabbit for gold grain
Body, therefore rabbit anteserum being capable of other epitopes of non-specific binding well.
The Selection experiment of reinforcing agent in 4 sample pad treatment fluid of test example
The color of different urines is multifarious, some Transparent color, some yellow, has plenty of honey color, and colloidal gold chromatographic
In the process true qualities be white nitrocellulose filter, when have it is out-of-date compared with dark colour sample flow the color of background can be caused deeper,
Therefore eliminate the color of background by adding toner so that the background of T/C lines becomes apparent from understanding, very clear, indirectly
Add sensitivity.
Liquid based on preparation pH9.0,0.1M Tris-HCL buffer solutions, adds hydroxypropyl methyl cellulose 0.1%,
PAA0.1%, 0.09M Na2CO3, rabbit anteserum 1%, then various concentration S9 mixings are coated on glass fibre element film, and coating is dense
It spends for 40ul/cm2, 25 DEG C are placed in, sample pad is made in humidity 10%~30%, drying and processing 18-22h.
Wherein, S9 is a kind of surfactant, and trade name is Tetronic 1307.Compared with other surfaces activity
Agent (such as S17 etc.) can bring the higher negative effect of false positive, and the addition of S9 does not interfere with testing result.
The spraying of gold grain and the preparation method of nitrocellulose membrane are the same as embodiment 1.
The preparation method of test strips is the same as embodiment 1.
The test strips that this test example is prepared are tested, and test method is the same as test example 1, test result such as 4 institute of table
Show.
The comparison that 4 reinforcing agent of table influences sample
Note：Totally 10 colour developing gradients：C1~C9, B.Wherein B represents " blank ", no band；C1~C9 represents that colour developing is strong
Degree, wherein number is bigger to represent that colour developing is more shallow.C8+ represents to develop the color between C8~C7, other and so on.In addition, in upper table
The numerical value representative before the % of S9 dosages is characterized compared with the buffer solution of every 100mL, the quality dosage (g) of S9.
From table 4, it can be seen that toner is similar to " bleaching ", the color of urine being carried in itself is eliminated into background
Color, improves the clarity of color, improves colour developing gradient indirectly.And low dosage effect is as high dose, therefore it is preferred that low
Dosage.
To sum up, cellulose family is as effective one kind in numerous auxiliary agents, and hydroxypropyl methyl cellulose contains numerous side chains,
Moisture can be absorbed to greatest extent and " expansion ", slow down the flowing of water, a liquid environment provided for the reaction of gold grain.
Its use scope is steadily fixed biomaterial and is not floated in 1%-10%, one stent of offer during spraying
Shifting is uniformly distributed, and ensure that the homogeneity of biomaterial.
K2CO3/Na2CO3One good alkaline environment is provided, maintains to add in the alkaline environment after urine sample jointly with Tris,
So as to eliminate the influence of false positive, and the dissolving rate of release of Immuno gold and film surface chromatography form are improved, even if pH meta-acids
Property urine (pH ＜ 7.0-7.5) gold grain can also be released effectively, expand detection pH applicabilities.
Tests below example 5~9 is the formulation screening test of the detection line coating buffer of the test strip of the present invention.
The preparation method screening of 5 gp160 solution of test example
1. urea is diluted to 1% respectively with Tris-HCl buffer solutions, 5%, 10% (wherein, 1% refers to 100mLTris-
1g ureas are added in HCl buffer solutions, the rest may be inferred by analogy), pH7.4,20mM of 400ul are then separately added into the 1st, 2,3,4 pipe
Tris-HCl；PH7.420mMTris-HCl+1% ureas；PH7.4 20mMTris-HCl+5% ureas；pH7.420mMTris-HCl+
10% urea then adds in gp160 into four pipes respectively, is diluted to 1.0mg/ml, the centrifugation of vortex mixing, Thermo
Nandrop 2000 measures the content of solution supernatant, calculates its dissolution rate.Dissolution rate calculation formula is as follows：
Dissolution rate=measurement concentration/theoretical concentration * 100%
5 different solvents of table dissolve the solubility of gp160
| Dilution pH9.0 | Tris-HCl | Tris-HCl+1% ureas | Tris-HCl+5% ureas | Tris-HCl+10% ureas |
| Liquid form | Flocculent deposit | Flocculent deposit | Supernatant liquid | Supernatant liquid |
| Dissolution rate | 0% | 30% | 98% | 98% |
As can be seen from Table 5, urea can be very good dissolving gp160 antigens, opens the hydrogen bond of water, makes the hydrophobic of albumen
Residue trails, and low concentration can only be partly dissolved, selection can all dissolve the minimum concentration of antigen with liquid, i.e.,
Tris-HCl+5% ureas.
The coating option screening of 6 coating protein of test example
The coating of coating protein is carried out according to three kinds of schemes of table 6, selects the hydrophilic nitrocellulose filters of milipore,
By film liquid amount 0.18ul/mm, by being sprayed onto on nitrocellulose filter for the coating protein fine uniform after coating, 25 DEG C are placed in, it is wet
Degree 10%~30%, drying and processing, 18-22h.
The method for coating screening of 6 coating protein of table
Sample pad is prepared by embodiment 1, and is diluted coating protein with coating solution according to the scheme of table 2, then is applied
It is distributed on nitrocellulose membrane, coating concentration 40ul/cm2.25 DEG C are dried 18-22h, humidity 10%-30%, for use.
Sample pad, nitrocellulose filter and blotting paper are sequentially pasted on bottom plate as described in Example 1, finished product is made
Test strips.
Selection Healthy People compares, the patient that makes a definite diagnosis of the detection HIV positives and with Healthy People feminine gender urine and the HIV positives
Strong sun, middle-jiao yang, function of the spleen and stomach, the internal reference of weak sun is made in Urine in Patients.2 drops (60-80ul) sample is added dropwise respectively to examination with rubber head dropper
In the sample pad of paper slip, due to capillarity, sample will be moved along test strips to sample pad and nitrocellulose filter, treat sample
Product by sample pad and nitrocellulose filter, as a result start to show completely；Observation shows result (note after 15 minutes：30 minutes
After develop the color it is invalid), the results are shown in Table 7.
Table 7 adds influences of the gp160 to sample sensitivity
Note：Totally 10 colour developing gradients：C1~C9, B.Wherein B represents " blank ", no band；C1~C9 represents that colour developing is strong
Degree, wherein number is bigger to represent that colour developing is more shallow.
As can be seen from Table 7, the weak positive sample (colour developing C8-C9) among scheme 2 may cause antibody due to the presence of urea
Capture ability decline, for strong, middle-jiao yang, function of the spleen and stomach sample almost without influence.The addition of gp160 can substantially enhance sample in scheme 3
Colour developing, improves sensitivity, and due to antigen when middle and high concentration urea has certain inhibitory action, colour developing is promoted limited.
Test example 7 eliminates the influence experiment of urea
For a small amount of urea on testing result without influence, the presence of excessive urea may influence the knot of antigen and antibody in body
It closes, finds that by adding a certain amount of S9 the influence of urea can be inhibited.By the use of pH7.420mMTris-HCl as matrix liquid, pass through
The gradient test of S9 has selected preferable 0.1% concentration (the Tris-HCl buffer solutions addition 0.1g S9 i.e. per 100mL)
Comparison is as follows：
Table 8 eliminates influence result of the urea to sample
Note：Totally 10 colour developing gradients：C1~C9, B.Wherein B represents " blank ", no band；C1~C9 represents that colour developing is strong
Degree, wherein number is bigger to represent that colour developing is more shallow.
Table 8 can be seen that urea has certain inhibition to colour developing, particularly weak positive sample；S9 causes colour developing more brilliant white；S9 and
The double solvents of urea eliminates the interference of urea well, while gold grain colour developing glossiness is better than individual gp41 controls effect；
When add in gp160 coating when, effectively capture the gp160 among urine caused by HIV antibody, compensate for gp41 correspondence
The missing inspection situation of antibody, sensitivity are effectively promoted, and rise 0.5-1 gradient so that the reference material of weak sun is by indistinctly difficult
The aubergine visually confirmed so that band is judged to develop the color.S9 extra simultaneously can enhance the glossiness of gold grain, visually
Improve color developing effect indirectly.
Influence experiment of the 8 gp160 concentration gradients of test example to sample
By the use of pH7.420mMTris-HCl as matrix liquid, influence of the different gp160 concentration to sample is test, as a result such as table 9
It is shown.
Influence result of the 9 gp160 concentration gradients of table to sample
Note：Totally 10 colour developing gradients：C1~C9, B.Wherein B represents " blank ", no band；C1~C9 represents that colour developing is strong
Degree, wherein number is bigger to represent that colour developing is more shallow.
Table 9 as can be seen that gp160 concentration in low concentration existing promotion effect, although high concentration sample is in part sample
There is the colour developing better than low concentration in this, but sample develops the color in naked eyes visible range, from cost consideration, preferably 0.5mg/
ml。
Not influence experiment of the isolabeling to sample of test example 9
Use K2CO3Aurosol particle is adjusted in the range of pH7.0-7.5, mouse IgG is added in and colloid gold particle albumen is (minimum
Dosage is 12 μ g/ml) mark forms colloid gold label mouse IgG antibody probe, and supernatant is removed in centrifugation, adds in 10ml 1%BSA and carries out
It is centrifuged again after closing and washs resuspension with Tris-HCl.
The dissolving of gp160, in the range of pH7.0-7.5, gp160 antigens and colloidal gold are carried out according to the method for test example 5
Granule protein (minimum amount is 12 μ g/ml) mark forms colloid gold label HIV specific antigen probes.Supernatant is removed in centrifugation, adds
Enter 10ml 1%BSA and carry out the piping and druming mixing that suspends, the suspended dispersed of particle is carried out using Ultrasonic Cell Disruptor, total ultrasonic time is about
2min, centrifugation go supernatant to wash resuspension with Tris-HCl.
Use K2CO3In the range of the pH8.0-8.5 for adjusting aurosol particle, gp36 and colloid gold particle albumen (minimum use are added in
Measure as 12 μ g/ml) mark forms colloid gold label gp36 probes, and supernatant is removed in centrifugation, adds in after 10ml1%BSA is closed again
Secondary centrifugation washs resuspension with Tris-HCl.
3 kinds of gold grains are concentrated into mixing respectively, with gold spraying instrument even application in sample pad, metal spraying width 6mm, length
28cm, sample pad length 2.8cm.25 DEG C are dried 18-22h, humidity 10%~30%, for use.
Not influence of the isolabeling to detection sample of table 10
Note：Totally 10 colour developing gradients：C1~C9, B.Wherein B represents " blank ", no band；C1~C9 represents that colour developing is strong
Degree, wherein number is bigger to represent that colour developing is more shallow.
The antibody that table 10 can be seen that exclusive use mouse anti-human igg can effectively capture HIV antibody, enhance sensitivity,
Simultaneously because its nonspecific presence so that the vacation sun of B+ occurs in part negative sample.Specific good, the sensitivity of gp160
It is shallow for the sample detection colour developing of weak sun not as good as the antibody of nonspecific mouse anti-human igg, it influences to judge.Pass through debugging two
The ratio of person, reaches the equalization point of sensitivity and specificity, and gp160, the mixtures of antibodies of mouse anti-human igg prevent that negative appearance is false
Sun, while in turn ensure positive deeper colour developing gradient.
Tests below example 10~13 is the performance test of the test strip of the present invention.
The sensitivity test of the test strips of 10 present invention of test example
Selection Healthy People compares, and the patient that the HIV positives are made a definite diagnosis carries out in the market the blood testing test paper of FDA registrations
Item (comparison test strips 1), saliva test strip (comparison test strips 2) carry out control test, test strips more of the invention
Sensitivity and specificity.2 drops (60-80ul) sample is added dropwise respectively to the sample pad of test strips with rubber head dropper, due to capillary
Pipe acts on, and sample will move along test strips to label pad and nitrocellulose filter, treat sample be completely dissolved gold grain and to
Nitrocellulose filter moves, and as a result starts to show；Observation shows result (note after 15 minutes：It is invalid to develop the color after 30 minutes).
As a result as shown in table 11.
The sensitivity test of 11 test strips of table
Note：Totally 10 colour developing gradients：C1~C9, B.Wherein B represents " blank ", no band；C1~C9 represents that colour developing is strong
Degree, wherein number is bigger to represent that colour developing is more shallow.
As can be seen from Table 11, the test scope (reference material is diluted to 10000 times) of test strips of the invention is than saliva, blood
Liquid test strip (comparison test strips 1 and 2, reference material is diluted to 100 times) wide, 100 times of high sensitivity or more.Secondly, saliva
Liquid, blood testing test strips are sensitive to urine specimen, and all positive urines can not detect, and the HIV urines examination of the present invention
Paper slip can detect urine and blood simultaneously, illustrate the urine detection test strips of the present invention and can replace blood to a certain extent
Test strip.
The country of the test strips of 11 present invention of test example refers to looping test
Nat'l Pharmaceutical ＆ Biological Products Control Institute HIV urine antibody reference materials are tested using the test strips of the embodiment of the present invention 1
(urine Rapid test), as a result as shown in table 12.
12 country of table refers to looping test result
Table 12 the result shows that, using embodiment 4 reagent strip detection urine HIV antibody sensitivity for 100%, specificity
Reach more than 99.9%, meet the standard of Nat'l Pharmaceutical ＆ Biological Products Control Institute's HIV antibody detection.
The clinical sample test of the test strips of 12 present invention of test example
The HIV patient 101 made a definite diagnosis through westernblot is selected from certain disease prevention and control center of province and (makes a definite diagnosis standard
It is:Western blot are positive, and sample number is P1~P101), with the health of Guangzhou Wondfo Biotech. Co., Ltd.
(standard is human urine sample：Body is without any clinical symptoms, and free from infection contact history, infection history, sample number is WF-N1~WF-
N100) 100 compare, and are tested using the test strips of the embodiment of the present invention 1, as a result as shown in Tables 13 and 14.
13 clinical sample test result of table
Note：Totally 10 colour developing gradients：C1~C9, B.Wherein B represents " blank ", no band；C1~C9 represents that colour developing is strong
Degree, wherein number is bigger to represent that colour developing is more shallow."-" is expressed as feminine gender；"+" expression is diagnosed as the positive.
14 clinical sample test result summary sheet of table
Find out from the result of table 13 and table 14, the clinical sample test display of small lot：Sensitivity >=99%, specificity
>=99%, meet expected requirement.
The clinical sample tracking and testing of the test strips of 13 present invention of test example
Using the test strips (abbreviation test strips 1) and comparison test strips (abbreviation test strips 2) of the embodiment of the present invention 1 to 5
The people at highest risk of example HIV positive patients carries out the tracking of 2 months by a definite date, and the comparison window time, the results are shown in Table 5.To having a competition
The agent prescription and preparation method of paper slip are identical with the test strips of embodiment 1, only difference is that：Detection zone is only coated with
There are HIV specificity recombinant antigen gp41 and gp36, be not coated with HIV specificity recombinant antigens gp160.
The tracking and testing of 15 suspicious patient of table
Note：Totally 10 colour developing gradients：C1~C9, B.Wherein B represents " blank ", no band；C1~C9 represents that colour developing is strong
Degree, wherein number is bigger to represent that colour developing is more shallow."-" is expressed as feminine gender；"+" expression is diagnosed as the positive.
As can be seen from Table 15, the addition of gp160 is so that the window time of some patientss shortens one week or so
Time (a suspect 2,5), when the comparison test strips test of these samples is negative, test strips of the invention are but shown as indistinct
Band can give some promptings of a suspect or precautionary measures to reduce the danger of propagation；Or cause part sample by general
Logical indistinct band colour developing is more dark bands (a suspect 1,3,4), is controlled so that medical staff makes a definite diagnosis or takes early
It treats.Later stage makes a definite diagnosis through " goldstandard " westernblot shows that band occur in 5 sample gp160, gp41 bands, with the present invention's
Test strips are consistent, which has strongly suggested that the sensitivity of the test strips of the present invention is splendid.
Claims (10)
1. a kind of test strips for detecting HIV antibody in urine, which is characterized in that the test strips include bottom plate and are arranged at
Sample pad, nitrocellulose filter and blotting paper on bottom plate, the both ends of the nitrocellulose filter overlap the sample pad respectively
And blotting paper, in the sample pad the nearly nitrocellulose filter end be coated with the mouse anti-human igg of colloid gold label antibody and
The gp160 antigens of colloid gold label and formed gold mark layer, the nitrocellulose filter be equipped with coating HIV recombinant antigens gp41 and
The first detection line of HIV recombinant antigens gp160 and the control line for being coated with sheep anti-mouse antibody.
2. the test strips of HIV antibody in detection urine according to claim 1, which is characterized in that the nitrocellulose
It is additionally provided with the second detection line of coating HIV recombinant antigens gp36 on film, the nearly nitrocellulose filter end is also in the sample pad
It is coated with the gp36 antigens of colloid gold label.
3. the test strips of HIV antibody in detection urine according to claim 1, which is characterized in that the nearly institute of gold mark layer
The distance between one end of nitrocellulose filter and described first detection line are stated as 4~7.5mm.
4. the test strips of HIV antibody in detection urine according to claim 1, which is characterized in that the sample pad is separate
It is 20~25mm that the distance between layer is marked in one end of the nitrocellulose filter with the gold.
5. a kind of test strips for detecting HIV antibody in urine, which is characterized in that the test strips include bottom plate and sequentially set
Sample pad, gold-labelled pad, nitrocellulose filter and the blotting paper being placed on bottom plate, one end of the gold-labelled pad are overlaid on the sample
Under pad, the other end of the gold-labelled pad and one end of blotting paper are overlapped on respectively on the nitrocellulose filter, the gold
Mark pads the antibody of mouse anti-human igg for being coated with colloid gold label and the gp160 antigens of colloid gold label and forms gold mark layer, institute
State the first detection line and bag that nitrocellulose filter is equipped with coating HIV recombinant antigen gp41 and HIV recombinant antigens gp160
By the control line of sheep anti-mouse antibody.
6. the test strips of HIV antibody in detection urine according to claim 5, which is characterized in that the nitrocellulose
The second detection line of coating HIV recombinant antigens gp36 is additionally provided on film, colloid gold label is also coated in the gold-labelled pad
Gp36 antigens.
7. the test strips of HIV antibody in detection urine according to claim 5, which is characterized in that the nearly institute of gold mark layer
The distance between one end of nitrocellulose filter and described first detection line are stated as 4~7.5mm.
8. a kind of liquid detection cup, including cup body, test strip clamp plate and cover film, at least a portion of the test strip clamp plate is placed in cup
In vivo, it which is provided with test paper slot, which is characterized in that be placed in the test paper slot any one of claim 1~7
Detection urine in HIV antibody test strips, the cover film is covered in the test strips surface by test paper slot opening face.
9. liquid detection cup according to claim 8, which is characterized in that left between the test strip clamp plate and the cup body
Space, the test strip clamp plate extend outwardly perpendicular to the bottom of cup and are stretched out from the cup body, the top side of the cup body
Along 6~15mm of gold mark layer of the test strips of HIV antibody in the detection urine.
10. liquid detection cup according to claim 8 or claim 9, which is characterized in that the test paper slot is equipped with several, institute
It states and illicit drugs inspection test strips is also placed in test paper slot, the illicit drugs inspection test strips and HIV antibody in the detection urine
Test strips be positioned in different test paper slots.
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