CN111965349A - Composition for fluorescence immunochromatography detection and application of composition in HIV detection - Google Patents
Composition for fluorescence immunochromatography detection and application of composition in HIV detection Download PDFInfo
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- 239000000203 mixture Substances 0.000 title claims abstract description 33
- 238000001514 detection method Methods 0.000 title abstract description 39
- 238000003317 immunochromatography Methods 0.000 title abstract description 9
- 239000004005 microsphere Substances 0.000 claims abstract description 32
- 238000004587 chromatography analysis Methods 0.000 claims abstract description 28
- 102000007327 Protamines Human genes 0.000 claims abstract description 25
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- 229940048914 protamine Drugs 0.000 claims abstract description 25
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 12
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- 210000002966 serum Anatomy 0.000 claims abstract description 8
- 101000582927 Photorhabdus laumondii subsp. laumondii (strain DSM 15139 / CIP 105565 / TT01) Lectin A Proteins 0.000 claims abstract description 5
- 239000000243 solution Substances 0.000 claims description 31
- 108010062580 Concanavalin A Proteins 0.000 claims description 18
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- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 claims description 5
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- 235000021240 caseins Nutrition 0.000 claims description 4
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 claims description 4
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 claims description 4
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- 239000003755 preservative agent Substances 0.000 claims 2
- 230000002335 preservative effect Effects 0.000 claims 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims 1
- 229930006000 Sucrose Natural products 0.000 claims 1
- 229940098773 bovine serum albumin Drugs 0.000 claims 1
- 239000005720 sucrose Substances 0.000 claims 1
- 240000003049 Canavalia gladiata Species 0.000 abstract description 3
- 235000010518 Canavalia gladiata Nutrition 0.000 abstract description 3
- 238000012795 verification Methods 0.000 abstract description 2
- 239000000523 sample Substances 0.000 description 33
- 239000012528 membrane Substances 0.000 description 10
- 239000000427 antigen Substances 0.000 description 8
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- 108091007433 antigens Proteins 0.000 description 8
- 241000725303 Human immunodeficiency virus Species 0.000 description 6
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- 229920002678 cellulose Polymers 0.000 description 5
- 239000001913 cellulose Substances 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 229920001220 nitrocellulos Polymers 0.000 description 5
- 239000013074 reference sample Substances 0.000 description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 239000002250 absorbent Substances 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 150000003384 small molecules Chemical class 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 102000011632 Caseins Human genes 0.000 description 3
- 108010076119 Caseins Proteins 0.000 description 3
- WHNWPMSKXPGLAX-UHFFFAOYSA-N N-Vinyl-2-pyrrolidone Chemical compound C=CN1CCCC1=O WHNWPMSKXPGLAX-UHFFFAOYSA-N 0.000 description 3
- 239000000020 Nitrocellulose Substances 0.000 description 3
- 230000002745 absorbent Effects 0.000 description 3
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- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 3
- 238000003908 quality control method Methods 0.000 description 3
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- 208000030507 AIDS Diseases 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 239000012491 analyte Substances 0.000 description 2
- 239000012472 biological sample Substances 0.000 description 2
- 229920002301 cellulose acetate Polymers 0.000 description 2
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- 238000000576 coating method Methods 0.000 description 2
- 238000004132 cross linking Methods 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 2
- 229910000397 disodium phosphate Inorganic materials 0.000 description 2
- 230000005284 excitation Effects 0.000 description 2
- 239000000835 fiber Substances 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- 238000001215 fluorescent labelling Methods 0.000 description 2
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 2
- 238000010166 immunofluorescence Methods 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 229920003023 plastic Polymers 0.000 description 2
- 239000004033 plastic Substances 0.000 description 2
- 229920000728 polyester Polymers 0.000 description 2
- 238000002791 soaking Methods 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 2
- 229910000162 sodium phosphate Inorganic materials 0.000 description 2
- DKIDEFUBRARXTE-UHFFFAOYSA-N 3-mercaptopropanoic acid Chemical group OC(=O)CCS DKIDEFUBRARXTE-UHFFFAOYSA-N 0.000 description 1
- 102000004506 Blood Proteins Human genes 0.000 description 1
- 108010017384 Blood Proteins Proteins 0.000 description 1
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- 108010027044 HIV Core Protein p24 Proteins 0.000 description 1
- 101000840258 Homo sapiens Immunoglobulin J chain Proteins 0.000 description 1
- 108060003951 Immunoglobulin Proteins 0.000 description 1
- 102100029571 Immunoglobulin J chain Human genes 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 230000002421 anti-septic effect Effects 0.000 description 1
- 230000000890 antigenic effect Effects 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 239000002274 desiccant Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
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- 238000000295 emission spectrum Methods 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 238000001917 fluorescence detection Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 239000003365 glass fiber Substances 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 102000018358 immunoglobulin Human genes 0.000 description 1
- 229940072221 immunoglobulins Drugs 0.000 description 1
- 230000036046 immunoreaction Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
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- 239000000178 monomer Substances 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 239000013307 optical fiber Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 229940069328 povidone Drugs 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 239000010453 quartz Substances 0.000 description 1
- 239000013558 reference substance Substances 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000012085 test solution Substances 0.000 description 1
- 239000000273 veterinary drug Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
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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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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Hematology (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Urology & Nephrology (AREA)
- Food Science & Technology (AREA)
- Physics & Mathematics (AREA)
- AIDS & HIV (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
A composition for fluorescence immunochromatography detection and application thereof in HIV detection. A composition for fluorescent microsphere chromatography test paper comprises Canavalia gladiata lectin A and protamine, and the dosage of Canavalia gladiata lectin A is 0.1g/cm2～1g/cm2The amount of protamine is 0.05g/cm2～0.15μg/cm2. The composition for the fluorescent microsphere chromatography test paper is applied to the fluorescent microsphere chromatography test paper and used for detecting HIV antibody positive serum, and proved by verification, the composition provided by the invention can obviously improve the sensitivity of a fluorescent chromatography reagent.
Description
Technical Field
The invention relates to a composition, in particular to a composition for a fluorescence detection reagent, which is used for improving the precision and the accuracy of antibody detection by the reagent.
Background
The fluorescence immunochromatography technology is a novel membrane detection technology based on antigen-antibody specific immunoreaction. The technology takes strip-shaped fiber chromatography materials fixed with a detection line (coated antibody or coated antigen) and a quality control line (anti-antibody) as a stationary phase, a test solution as a mobile phase, a fluorescence labeled antibody or antigen fixed on a connecting pad, and an analyte to be analyzed moves on the chromatography strip through capillary action. For macromolecular antigens (proteins, viruses, pathogenic bacteria and the like) with a plurality of antigenic determinants, a sandwich-type double-antibody sandwich immunochromatography method is generally adopted, namely, an object to be detected is firstly combined with a fluorescence labeling antibody under the action of a mobile phase, and then is combined with a coating antibody to form a sandwich-type double-antibody sandwich when reaching a detection line. For small molecule antigens (veterinary drugs, prohibited drugs and the like) with only a single epitope, after the small molecule antigens to be detected are combined with the fluorescence labeling antibody, the small molecule antigens are difficult to be combined with the coating antibody on the detection line due to steric hindrance. Therefore, the small molecule analyte with single epitope is mostly detected by using the competitive immunochromatography.
The immunochromatographic test strip generally comprises a filter pad, a sample pad, a conjugate pad, a reaction pad, a water-absorbent pad, and the like. The filter pad is a quartz fiber filter paper, and can filter large-particle substances in blood and avoid interference on a detection result. The sample pad is a polyester cellulose film or a nitrocellulose film or a cellulose acetate film, and before the test paper is assembled, the sample pad needs to be treated by using a sample pad treatment solution, and the sample pad is dried at 37 ℃ and then used. The combination pad is a polyester cellulose film or a nitrocellulose film or a cellulose acetate film, and before the test paper is assembled, the combination pad needs to be treated by using a combination pad treatment solution, and the combination pad is dried at 37 ℃ and then used for adsorbing the fluorescent microsphere marker. The reaction pad is a nitrocellulose membrane (also called NC membrane) for binding an antibody, and as a detection area of an experimental result, a detection line (also called T line, i.e., for determining whether a substance to be detected exists in a sample) and a quality control line (also called C line, i.e., for determining whether the test paper can achieve the purpose of detection to prevent the T line from being missed) are usually provided. In addition, the absorbent pad is a filter paper material having uniform water absorbency, and is disposed at the extreme end of the entire chromatography to provide power for the chromatography reaction while inhibiting reflux.
The patent application CN200910142348.3 discloses a fluorescent microsphere immunochromatography detection card for quantitatively detecting AIDS virus, which adopts an immunochromatography technology to realize quantitative immunoassay of the AIDS virus. In the detection process, a fluorescent microsphere excitation light source is adopted for excitation, all emission spectra are collected, gathered and multiplied by a CCD scanning technology or an optical fiber technology after emitted fluorescence passes through an optical filter device, and then are converted into numerical signals, and the concentration of an object to be detected is automatically calculated by using built-in analysis software in a fluorescence analyzer. The detection card comprises A, B test strips, a sample pad, a glass fiber membrane, a nitrocellulose membrane and absorbent paper, wherein a detection line and a quality control line are fixed on the nitrocellulose membrane to realize the simultaneous detection of HIV antibody and HIV P24 antigen. The invention has the advantages of high sensitivity, accurate quantification, rapid detection, convenient operation, economy and practicality.
Patent application CN201510542660.7 discloses a confining liquid for reducing false positive of biological sample in vitro detection, which can reduce nonspecific adsorption of serum protein and reduce substrate effect of serum, thereby reducing false positive of biological sample in vitro detection, improving detection accuracy, and having high detection sensitivity, wide detection linearity and strong stability. The confining liquid comprises a reagent A and a reagent B, wherein the reagent A is an amino acid solution, and the reagent B is a 3-mercaptopropionic acid solution. The confining liquid is used for confining antibody microspheres in an immunofluorescence chromatography detection system. The specific application process is as follows: pretreating the fluorescent microspheres, then crosslinking the fluorescent microspheres with antibodies to obtain antibody microspheres, sealing the antibody microspheres with a sealing solution, and finally storing the antibody microspheres with a buffer solution. The ionic amino acid monolayer obtained after the reaction of the two reagents in the confining liquid is distributed on the surface of the antibody microsphere in the immunofluorescence chromatography detection system.
The total antibody detection kit can detect IgG and IgM simultaneously, and IgM in a pentamer form easily causes aggregation and crosslinking of fluorescent microspheres, so that the collection of fluorescent signals is influenced, and the detection precision and accuracy are not ideal. The IgM antibodies in the serum/plasma sample account for 5-10% of the total amount of serum immunoglobulins, and the serum concentration is about 1 mg/mL. The pentameric form of IgM is composed of 5 monomeric subunits linked by disulfide bonds to a J chain, which is a cyclic chain linking five subunits. The heavy and light chains of each monomer molecule are linked by disulfide bonds. The IgM pentamer has 10 antigen binding fragments (Fab), can simultaneously bind to 10 antigens, and is easier to generate a cross-linked structure, so that fluorescent microspheres are gathered on a chromatographic membrane of a chromatographic system to influence signal acquisition.
Disclosure of Invention
An object of the present invention is to provide a composition for use in fluorescence immunochromatography detection, which improves the sensitivity of detection.
The invention also aims to provide a composition for manufacturing the fluorescent microsphere chromatography test paper, so that the detection sensitivity is improved.
It is still another object of the present invention to provide a composition, which is applied to a sample pad and a filter pad of a fluorescent microsphere chromatography test paper to improve detection sensitivity.
Still another object of the present invention is to provide the use of a composition for preparing a kit for detecting HIV antibody positive sera, which is advantageous for improving the sensitivity of the detection.
The fifth purpose of the present invention is to provide a fluorescent microsphere chromatography test paper, wherein the sample pad and the filter pad of the fluorescent microsphere chromatography test paper respectively adsorb the composition, so as to improve the detection sensitivity.
A composition for fluorescent microsphere chromatography test paper comprises concanavalin A and protamine, wherein the dosage of concanavalin A is 0.1g/cm2～1g/cm2The amount of protamine is 0.05g/cm2～0.15μg/cm2。
The other composition for the fluorescent microsphere chromatography test paper comprises a sample pad treatment solution and a filter pad treatment solution, wherein the sample pad treatment solution comprises protamine, and the dosage of the protamine is 0.05g/cm2～0.15μg/cm2(ii) a The filter pad treatment solution comprises concanavalin A in an amount of 0.1g/cm2～1g/cm2。
The other composition for the fluorescent microsphere chromatography test paper comprises a sample pad treatment solution and a filter pad treatment solution, wherein the sample pad treatment solution comprises phosphate buffered saline (pH7.2), Casein (Casein), Tween-20, povidone (PVP K-30) and protamine, and the adsorption amount of the protamine on the sample pad is 0.05g/cm2～0.15μg/cm2(ii) a The filter pad treatment solution comprises phosphate-buffered saline (pH7.2), emulsifier (such as Pluracare 1307), antiseptic (such as Proclin-300), and Canavalia gladiata lectin A, which is adsorbed on the filter pad in an amount of 0.1g/cm2～1g/cm2。
Another composition for use in a fluorescent microsphere chromatography strip, comprising:
the sample pad treatment solution contains protamine with the concentration of 1 g/L;
a filter pad treatment liquid containing concanavalin A at a concentration of 5g/L, and
the test strip comprises a sample pad and a filter pad;
the sample pad was wetted with the sample pad treatment solution so that the amount of protamine adsorbed on the sample pad was 0.05g/cm2～0.15μg/cm2Wetting the filter pad with a filter pad treatment solution so that the amount of concanavalin A adsorbed on the filter pad is 0.1g/cm2～1g/cm2。
The composition provided by the invention is used for detecting HIV antibody positive serum, and the composition provided by the invention can obviously improve the sensitivity of a fluorescence chromatography reagent by verification.
A fluorescence chromatography test paper comprises a sample pad and a filter pad, wherein the adsorption amount of protamine on the sample pad is 0.05g/cm2～0.15μg/cm2(ii) a The adsorption amount of Canavalia ensiformis lectin A on the filter pad was 0.1g/cm2～1g/cm2。
Detailed Description
The technical solution of the present invention is described in detail below. Although the present invention has been described in detail with reference to the preferred embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the spirit and scope of the invention as defined in the appended claims.
Example 1
Reagents such as a filter pad treatment solution, a sample pad treatment solution, and a conjugate pad treatment solution were prepared as shown in tables 1, 2, and 3 below.
TABLE 1
| NaH2PO4·2H2O | 0.6g/L |
| Na2HPO4 | 2.3g/L |
| NaCl | 8.5g/L |
| Concanavalin A | 5g/L |
| Pluracare 1307 | 5g/L |
| Proclin-300 | 0.1g/L |
TABLE 2
| NaH2PO4·2H2O | 0.6g/L |
| Na2HPO4 | 2.3g/L |
| NaCl | 8.5g/L |
| Casein protein | 5g/L |
| PVP K-30 | 5g/L |
| Tween-20 | 5g/L |
| Protamine | 1g/L |
TABLE 3
Adding the prepared sample pad treatment solution (filter pad treatment solution or combined pad treatment solution) into a plastic box, putting the glass cellulose membranes into the plastic box one by one, completely soaking the glass cellulose membranes in the treatment solution, taking out after soaking for 10 minutes, draining, putting on a plate frame, and putting in an oven at 37 ℃ for 12 hours to obtain the glass cellulose membrane. Putting it into a self-sealing bag filled with a drying agent for later use. The amount of protamine adsorbed by the prepared sample pad was 0.05g/cm2～0.15μg/cm2The amount of concanavalin A adsorbed on the obtained filter pad was 0.1g/cm2～1g/cm2。
The filter pad, the sample pad, the conjugate pad, the reaction pad, and the absorbent pad are sequentially disposed to prepare the immunochromatographic test strip of this embodiment.
Separately, an immunochromatographic test strip containing no protamine and concanavalin A and an immunochromatographic test strip containing only protamine or concanavalin A were prepared according to the above-mentioned methods.
Example 2
Taking a strong positive sample P1, diluting the strong positive sample step by step according to sesqui, and preparing positive test products which are respectively marked as P2-P10.
And taking the negative standard substance N1, diluting the negative standard substance by times, and preparing negative reference substances which are respectively marked as N2-N10.
The above positive test samples and negative reference samples were tested with the Yapei 4 th generation AIDS detection paper, and the results are shown in tables 4 and 5, respectively.
TABLE 4
| P1 | P2 | P3 | P4 | P5 | P6 | P7 | P8 | P9 | P10 |
| + | + | + | + | + | + | + | + | + | - |
TABLE 5
| N1 | N2 | N3 | N4 | N5 | N6 | N7 | N8 | N9 | N10 |
| - | - | - | - | - | - | - | - | - | - |
As can be seen from tables 4 and 5, the negative reference products showed no positive results, whereas 1 of the positive test products showed negative results, which are attributed to multiple sesquidilutions exceeding the detection limit of the reagents.
Example 3
The immunochromatographic test strip containing no protamine and concanavalin a, the immunochromatographic test strip containing concanavalin a only, the immunochromatographic test strip containing protamine only, and the test strip prepared in this example detected each positive test sample and each negative reference sample in example 2, and measured with an AFS-1000 type fluorescence detector (purchased from tibo biotechnology limited, guangzhou), the maximum absorption peak area pattern was selected, read and recorded, and the CUTOFF value was 500. See tables 6, 7, 8 and 9, respectively, for the results.
TABLE 6
As can be seen in table 6, P10 showed a negative result in the positive test sample, while N1, N5 and N8 all showed positive results in the negative reference sample.
TABLE 7
As can be seen from table 7, P9 and P10 in the positive test sample showed negative results and N5 in the negative reference sample showed positive results when tested using an immunochromatographic strip containing only concanavalin a.
TABLE 8
As can be seen in Table 8, P10 showed a negative result in the positive test sample, while N1 and N5 showed positive results in the negative reference sample, tested using the immunochromatographic strip containing protamine only.
TABLE 9
As can be seen in table 9, when the immunochromatographic test strip containing both protamine and concanavalin a was used for the test, P10 showed a negative result in the positive test sample, whereas no positive result was observed in the negative reference sample. The results are consistent with those of the 4 th generation AIDS detection paper of Yapei.
Claims (12)
1. A composition for fluorescent microsphere chromatography test paper is characterized by comprising concanavalin A and protamine.
2. The composition for fluorescent microsphere chromatographic test paper according to claim 1, wherein the concanavalin A is used in an amount of 0.1g/cm2～1g/cm2。
3. The composition for fluorescent microsphere chromatography test paper of claim 1, wherein the amount of protamine is 0.05g/cm2～0.15μg/cm2。
4. The composition of claim 1 for use in a fluorescent microsphere chromatographic test strip comprising
A sample pad treatment liquid containing protamine at a concentration of 1 g/L;
the filter pad treatment solution contained concanavalin A at a concentration of 5 g/L.
5. The composition for fluorescent microsphere chromatography test paper according to claim 1, characterized by comprising:
the sample pad treatment solution contains protamine with the concentration of 1 g/L;
a filter pad treatment liquid containing concanavalin A at a concentration of 5g/L, and
the test strip comprises a sample pad and a filter pad;
the sample pad was wetted with the sample pad treatment solution so that the amount of protamine adsorbed on the sample pad was 0.05g/cm2～0.15μg/cm2Wetting the filter pad with a filter pad treatment solution so that the amount of concanavalin A adsorbed on the filter pad is 0.1g/cm2～1g/cm2。
6. The composition of claim 5, wherein the sample pad treatment solution further comprises casein, PVP K-30, Tween-20 and phosphate buffered saline.
7. The composition for fluorescent microsphere chromatography test paper of claim 5, wherein said filter pad treatment solution further comprises Pluracare 1307, phosphate buffered saline and preservative Proclin-300.
8. The composition of claim 5, further comprising a treatment solution containing sucrose, casein, bovine serum albumin, PVP K-30, Tween-20, Proclin-300 as preservative, and phosphate buffered saline.
9. The composition for fluorescent microsphere chromatography test paper according to any one of claims 1 to 8, wherein the composition is used for detecting HIV antibody positive serum and improving the sensitivity of a fluorescent chromatography reagent.
10. Use of the composition for fluorescent microsphere chromatography test paper according to any one of claims 1 to 8 in the preparation of a kit for detecting HIV antibody positive serum.
11. A kit comprising the composition for fluorescent microsphere chromatography test paper according to any one of claims 1 to 8.
12. The fluorescent chromatographic test paper is characterized by comprising a sample pad and a filter pad, wherein the adsorption capacity of protamine on the sample pad is 0.05g/cm2～0.15μg/cm2(ii) a The adsorption amount of Canavalia ensiformis lectin A on the filter pad was 0.1g/cm2～1g/cm2。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911188396.6A CN111965349A (en) | 2019-11-28 | 2019-11-28 | Composition for fluorescence immunochromatography detection and application of composition in HIV detection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201911188396.6A CN111965349A (en) | 2019-11-28 | 2019-11-28 | Composition for fluorescence immunochromatography detection and application of composition in HIV detection |
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Citations (6)
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| US5558834A (en) * | 1991-10-03 | 1996-09-24 | Bayer Corporation | Device and method of seperating and assaying whole blood |
| US20020045195A1 (en) * | 2000-10-13 | 2002-04-18 | Hubscher Thomas T. | Method for the visual detection of specific antibodies in human serum by the use of lateral flow assays |
| US20060166374A1 (en) * | 2005-01-21 | 2006-07-27 | Hubscher Thomas T | Method for the visual detection of specific antibodies by the use of lateral flow assays |
| WO2016052690A1 (en) * | 2014-10-02 | 2016-04-07 | コニカミノルタ株式会社 | Blocking method for immunoassay, and immunoassay instrument |
| CN106814193A (en) * | 2016-12-23 | 2017-06-09 | 美康生物科技股份有限公司 | Neutrophil gelatinase-associated lipocalin reagent box for detecting content |
| CN107144693A (en) * | 2017-05-02 | 2017-09-08 | 暨南大学 | Detect CD4 in blood+Lateral chromatography kit of T cell quantity and preparation method thereof |
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| US5558834A (en) * | 1991-10-03 | 1996-09-24 | Bayer Corporation | Device and method of seperating and assaying whole blood |
| US20020045195A1 (en) * | 2000-10-13 | 2002-04-18 | Hubscher Thomas T. | Method for the visual detection of specific antibodies in human serum by the use of lateral flow assays |
| US20060166374A1 (en) * | 2005-01-21 | 2006-07-27 | Hubscher Thomas T | Method for the visual detection of specific antibodies by the use of lateral flow assays |
| WO2016052690A1 (en) * | 2014-10-02 | 2016-04-07 | コニカミノルタ株式会社 | Blocking method for immunoassay, and immunoassay instrument |
| CN106814193A (en) * | 2016-12-23 | 2017-06-09 | 美康生物科技股份有限公司 | Neutrophil gelatinase-associated lipocalin reagent box for detecting content |
| CN107144693A (en) * | 2017-05-02 | 2017-09-08 | 暨南大学 | Detect CD4 in blood+Lateral chromatography kit of T cell quantity and preparation method thereof |
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