CN115856326A - 25 hydroxy vitamin D detect reagent box - Google Patents
25 hydroxy vitamin D detect reagent box Download PDFInfo
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- CN115856326A CN115856326A CN202310185323.1A CN202310185323A CN115856326A CN 115856326 A CN115856326 A CN 115856326A CN 202310185323 A CN202310185323 A CN 202310185323A CN 115856326 A CN115856326 A CN 115856326A
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- JWUBBDSIWDLEOM-XHQRYOPUSA-N (3e)-3-[(2e)-2-[1-(6-hydroxy-6-methylheptan-2-yl)-7a-methyl-2,3,3a,5,6,7-hexahydro-1h-inden-4-ylidene]ethylidene]-4-methylidenecyclohexan-1-ol Chemical compound C1CCC2(C)C(C(CCCC(C)(C)O)C)CCC2\C1=C\C=C1/CC(O)CCC1=C JWUBBDSIWDLEOM-XHQRYOPUSA-N 0.000 title claims abstract description 58
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- 238000006243 chemical reaction Methods 0.000 claims abstract description 47
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- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims description 6
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- QYSXJUFSXHHAJI-XFEUOLMDSA-N Vitamin D3 Natural products C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C/C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-XFEUOLMDSA-N 0.000 claims description 5
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Abstract
The invention relates to a 25 hydroxy vitamin D detection kit, which relates to the technical field of in vitro diagnosis and comprises a test strip, wherein the test strip comprises a PVC plate, the PVC plate sequentially comprises a water absorption area, a reaction area, a connecting bridge, a release area and a sample application area from left to right, and the connecting bridge is one of polyurethane, glass fiber, cellulose and polyester films.
Description
Technical Field
The invention relates to the technical field of in-vitro diagnosis, in particular to a 25-hydroxy vitamin D detection kit.
Background
As early as the 30 s of the 20 th century, scientists discovered that much sun exposure or edible UV-irradiated olive oil, linseed oil, etc. could resist chondropathy, and named the active ingredient vitamin D. Vitamin D is a necessary nutrient for human and livestock, poultry growth, breeding, life support and health maintenance.
Vitamin D obtained from both dietary and skin pathways is transported to the liver in association with plasma alpha-globulin, and first becomes 25-hydroxyvitamin D in the endoplasmic reticulum and mitochondria of hepatocytes by the action of 25-hydroxylase. The high or low level of serum 25-hydroxyvitamin D may reflect the storage level of vitamin D in humans and is associated with clinical symptoms of vitamin D deficiency.
At present, the commonly used detection methods of 25-hydroxy vitamin D comprise a chemiluminescence method, an enzyme-linked immunosorbent assay, an electrochemiluminescence method and the like, most of the detection methods are long in time consumption and complex in operation, and large-scale instruments and equipment are needed, so that the detection is not beneficial to realizing generalization. Compared with the prior art, the immunochromatography method is simple, convenient and quick to operate, does not need additional instruments and equipment, can realize detection basification, and is convenient to use in primary hospitals, clinics and remote areas.
Disclosure of Invention
The invention aims to provide a 25-hydroxy vitamin D detection kit, which has the advantages of convenience in carrying, simplicity in operation, high sensitivity and coincidence rate, high stability and the like, is suitable for home self-testing, primary clinics and medical institutions without large-scale instruments and equipment, is beneficial to realizing socialization and generalization detection, and improves the physical quality of people.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows: the utility model provides a 25 hydroxyvitamin D detect reagent box, includes the test paper strip, and the test paper strip includes the PVC board, from left to right includes water absorption area, reaction zone, connecting bridge, release district, adds the appearance district in proper order on the PVC board, and the connecting bridge is one of polyurethane, glass fiber, cellulose, polyester film.
As an optimization, the connecting bridge is cellulose.
As an optimized scheme, the preparation method of the connecting bridge comprises the following steps: soaking the cellulose in 40mL of treatment solution, standing for 24h, drying at 35 ℃ for 12h, and cutting to the required width.
As an optimization scheme, the treatment solution comprises 20-50mM of PBS buffer solution, 0.2-1% of BSA, 0.5-1.2% of PVP, 0.2-1% of Tween and 0.2-1.5% of Triton.
As an optimization scheme, the treatment solution comprises 30mM of PBS buffer solution, 0.6% of BSA, 0.5% of PVP, 0.5% of Tween and 0.8% of triton.
As an optimization, the length of the connecting bridge is 6mm.
As an optimized scheme, one end of the water absorption area is lapped on one end of the reaction area, one end of the connecting bridge is lapped on the other end of the reaction area, one end of the release area is lapped on the other end of the connecting bridge, and one end of the sample addition area is lapped on the other end of the release area.
As an optimized scheme, the lapping width of the connecting bridge and the reaction zone is 0.7-1mm, and the lapping width of the release zone and the connecting bridge is 1mm.
As an optimized scheme, the water absorption area is water absorption paper, the reaction area is a nitrocellulose membrane coated with 25 hydroxy vitamin D antigen, the release area is a gold-labeled solid phase pad with a solid phase containing 25 hydroxy vitamin D antibody, and the sample adding area is a sample pad.
As an optimized scheme, the length of the water absorption area is 22mm, the length of the reaction area is 20mm, the length of the release area is 5mm, the length of the sample adding area is 14mm, the overlapping width of the water absorption area and the reaction area is 1mm, and the overlapping width of the sample adding area and the release area is 1.5-2mm.
By adopting the technical scheme, the invention has the following advantages: the invention prolongs the reaction channel of the antigen and the antibody and increases the reaction time of the antigen and the antibody by adding the connecting bridge which is specially processed, thereby greatly improving the sensitivity of the detection reagent.
The invention is further described with reference to the following figures and detailed description.
Drawings
FIG. 1 is a schematic structural diagram of a test paper strip of a 25-hydroxyvitamin D detection kit according to the present invention;
fig. 2 is a schematic view of the structure of a test strip in a comparative example.
In the figure, the position of the upper end of the main shaft,
1-water absorption zone, 2-reaction zone, 3-connecting bridge, 4-release zone, 5-sample addition zone and 6-PVC plate.
Detailed Description
Embodiments of the present invention will be described in detail below with reference to examples, in which specific conditions are not specified, and which are performed according to conventional conditions or conditions recommended by manufacturers. The reagents or instruments used are not indicated by manufacturers, and are all conventional products available on the market.
The embodiments described are only a part of the embodiments of the present invention, and not all embodiments, and all other embodiments obtained by those skilled in the art based on the embodiments of the present invention without any inventive work belong to the protection scope of the present invention.
All percentages in the following examples are by weight.
Example 1
As shown in fig. 1, a 25 hydroxyvitamin D detection kit comprises a test strip, the test strip comprises a PVC plate 6, the PVC plate 6 sequentially comprises a water absorption region 1, a reaction region 2, a connecting bridge 3, a release region 4, and a sample addition region 5 from left to right, one end of the water absorption region 1 is lapped on one end of the reaction region 2, one end of the connecting bridge 3 is lapped on the other end of the reaction region 2, one end of the release region 4 is lapped on the other end of the connecting bridge 3, one end of the sample addition region 5 is lapped on the other end of the release region 4, the water absorption region 1 is absorbent paper, the reaction region 2 is a nitrocellulose membrane coated with 25 hydroxyvitamin D antigen, the release region 4 is a gold-labeled solid phase pad with 25 hydroxyvitamin D antibody in solid phase, the sample addition region 5 is a sample pad, and the connecting bridge 3 is polyurethane. The length of connecting bridge 3 is 6mm, the length of absorbing water district 1 is 22mm, the length of reaction zone 2 is 20mm, the length of release area 4 is 5mm, the length of application of sample district 5 is 14mm, the overlap joint width of absorbing water district 1 and reaction zone 2 is 1mm, the overlap joint width of connecting bridge 3 and reaction zone 2 is 0.7-1mm, the overlap joint width of release area 4 and connecting bridge 3 is 1mm, the overlap joint width of application of sample district 5 and release area 4 is 1.5-2mm.
Example 2
As shown in fig. 1, a 25 hydroxyvitamin D detection kit comprises a test strip, the test strip comprises a PVC plate 6, the PVC plate 6 sequentially comprises a water absorption region 1, a reaction region 2, a connecting bridge 3, a release region 4, and a sample addition region 5 from left to right, one end of the water absorption region 1 is lapped on one end of the reaction region 2, one end of the connecting bridge 3 is lapped on the other end of the reaction region 2, one end of the release region 4 is lapped on the other end of the connecting bridge 3, one end of the sample addition region 5 is lapped on the other end of the release region 4, the water absorption region 1 is absorbent paper, the reaction region 2 is a nitrocellulose membrane coated with 25 hydroxyvitamin D antigen, the release region 4 is a gold-labeled solid phase pad with 25 hydroxyvitamin D antibody in solid phase, the sample addition region 5 is a sample pad, and the connecting bridge 3 is glass fiber. The length of the connecting bridge 3 is 6mm, the length of the water absorption area 1 is 22mm, the length of the reaction area 2 is 20mm, the length of the release area 4 is 5mm, the length of the sample addition area 5 is 14mm, the lap joint width of the water absorption area 1 and the reaction area 2 is 1mm, the lap joint width of the connecting bridge 3 and the reaction area 2 is 0.7-1mm, the lap joint width of the release area 4 and the connecting bridge 3 is 1mm, and the lap joint width of the sample addition area 5 and the release area 4 is 1.5-2mm.
Example 3
As shown in figure 1, a 25 hydroxyvitamin D detection kit comprises a test strip, the test strip comprises a PVC plate 6, the PVC plate 6 sequentially comprises a water absorption area 1, a reaction area 2, a connecting bridge 3, a release area 4 and a sample adding area 5 from left to right, one end of the water absorption area 1 is lapped on one end of the reaction area 2, one end of the connecting bridge 3 is lapped on the other end of the reaction area 2, one end of the release area 4 is lapped on the other end of the connecting bridge 3, one end of the sample adding area 5 is lapped on the other end of the release area 4, the water absorption area 1 is absorbent paper, the reaction area 2 is a nitrocellulose membrane coated with 25 hydroxyvitamin D antigen, the release area 4 is a gold-labeled solid phase pad with 25 hydroxyvitamin D antibody in solid phase, the sample adding area 5 is a sample pad, and the connecting bridge 3 is cellulose. The length of connecting bridge 3 is 6mm, the length of absorbing water district 1 is 22mm, the length of reaction zone 2 is 20mm, the length of release area 4 is 5mm, the length of application of sample district 5 is 14mm, the overlap joint width of absorbing water district 1 and reaction zone 2 is 1mm, the overlap joint width of connecting bridge 3 and reaction zone 2 is 0.7-1mm, the overlap joint width of release area 4 and connecting bridge 3 is 1mm, the overlap joint width of application of sample district 5 and release area 4 is 1.5-2mm.
Example 4
As shown in fig. 1, a 25 hydroxyvitamin D detection kit comprises a test strip, the test strip comprises a PVC plate 6, the PVC plate 6 sequentially comprises a water absorption region 1, a reaction region 2, a connecting bridge 3, a release region 4, and a sample addition region 5 from left to right, one end of the water absorption region 1 is lapped on one end of the reaction region 2, one end of the connecting bridge 3 is lapped on the other end of the reaction region 2, one end of the release region 4 is lapped on the other end of the connecting bridge 3, one end of the sample addition region 5 is lapped on the other end of the release region 4, the water absorption region 1 is water absorption paper, the reaction region 2 is a nitrocellulose membrane coated with 25 hydroxyvitamin D antigen, the release region 4 is a gold-labeled solid phase pad with 25 hydroxyvitamin D antibody in solid phase, the sample addition region 5 is a sample pad, and the connecting bridge 3 is a polyester membrane. The length of connecting bridge 3 is 6mm, the length of absorbing water district 1 is 22mm, the length of reaction zone 2 is 20mm, the length of release area 4 is 5mm, the length of application of sample district 5 is 14mm, the overlap joint width of absorbing water district 1 and reaction zone 2 is 1mm, the overlap joint width of connecting bridge 3 and reaction zone 2 is 0.7-1mm, the overlap joint width of release area 4 and connecting bridge 3 is 1mm, the overlap joint width of application of sample district 5 and release area 4 is 1.5-2mm.
Comparative example: as shown in FIG. 2, the test strip comprises a PVC plate 6, and the PVC plate 6 sequentially comprises a water absorption region 1, a reaction region 2, a release region 4 and a sample addition region 5 from left to right.
20 serum samples were selected and the reagents were compared to control and examples 1-4 using Roche (25 hydroxy vitamin D assay kit, AC-57F 1) reagent.
The results show that the sensitivity of the examples 1 to 4 is obviously improved compared with that of the comparative example, wherein the sensitivity of the example 3, namely the connecting bridge 3, is most obviously improved when the cellulose is selected. Cellulose has the most compact pores, polyester film second, and glass fiber loosest. The small pore folded structure has lower permeability and lower permeation rate, and more target analyte interacts with the antibody on the conjugate pad due to the stacking effect and longer interaction time caused by the small pores in the cellulose stacking pad, so that the signal enhancement of the test result can be observed.
Example 5
A preparation method of a 25-hydroxy vitamin D detection kit comprises a preparation method of a connecting bridge 3, wherein the preparation method of the connecting bridge 3 comprises the following steps: soaking 25 × 30cm cellulose in 40mL of the treatment solution, standing for 24h, drying at 35 deg.C for 12h, and cutting to desired width. The treatment solution comprises 20mM of PBS buffer solution, 0.5% of BSA, 0.7% of PVP, 0.5% of Tween and 0.5% of triton.
The PBS buffer solution has the buffer function of adjusting proper pH, and can fully ensure the biological activity; BSA generally acts as a stabilizer, acting as a "stabilizer" and a "carrier"; PVP, tween 20 and Triton are used as surfactants, so that the PVP, tween 20 and Triton have good non-dissociation property, play a certain role in helping dissolution, can inhibit the automatic polymerization of macromolecules and improve the stability.
Example 6
A preparation method of a 25-hydroxy vitamin D detection kit comprises a preparation method of a connecting bridge 3, wherein the preparation method of the connecting bridge 3 comprises the following steps: soaking 25 × 30cm cellulose in 40mL of the treatment solution, standing for 24h, drying at 35 deg.C for 12h, and cutting to desired width. The treatment solution comprises 30mM of PBS buffer solution, 0.5% of BSA, 0.7% of PVP, 0.5% of Tween and 0.5% of triton.
The PBS buffer solution has the buffer function of adjusting proper pH, and can fully ensure the biological activity; BSA generally acts as a stabilizer, acting as a "stabilizer" and a "carrier"; PVP, tween 20 and Triton are used as surfactants, so that the PVP, tween 20 and Triton have good non-dissociation property, play a certain role in helping dissolution, can inhibit the automatic polymerization of macromolecules and improve the stability.
Example 7
A preparation method of a 25-hydroxy vitamin D detection kit comprises a preparation method of a connecting bridge 3, wherein the preparation method of the connecting bridge 3 comprises the following steps: soaking 25 × 30cm cellulose in 40mL of the treatment solution, standing for 24h, drying at 35 deg.C for 12h, and cutting to desired width. The treatment solution comprises 40mM of PBS buffer solution, 0.5% of BSA, 0.7% of PVP, 0.5% of Tween and 0.5% of triton.
The PBS buffer solution has the buffer function of adjusting proper pH, and can fully ensure the biological activity; BSA generally acts as a stabilizer, acting as a "stabilizer" and a "carrier"; PVP, tween 20 and Triton are used as surfactants, so that the PVP, tween 20 and Triton are good in non-dissociative property, a certain solubilizing effect is achieved, the autopolymerization of macromolecules can be inhibited, and the stability is improved.
Example 8
The preparation method of the 25-hydroxy vitamin D detection kit comprises a preparation method of a connecting bridge 3, wherein the preparation method of the connecting bridge 3 comprises the following steps: soaking 25 × 30cm cellulose in 40mL of the treatment solution, standing for 24h, drying at 35 deg.C for 12h, and cutting to the desired width. The treatment solution comprises 50mM of PBS buffer solution, 0.5% of BSA, 0.7% of PVP, 0.5% of Tween and 0.5% of triton.
The PBS buffer solution has the buffer function of adjusting proper pH, and can fully ensure the biological activity; BSA generally acts as a stabilizer, acting as a "stabilizer" and a "carrier"; PVP, tween 20 and Triton are used as surfactants, so that the PVP, tween 20 and Triton have good non-dissociation property, play a certain role in helping dissolution, can inhibit the automatic polymerization of macromolecules and improve the stability.
Experiment 1: 20 serum samples were selected and the reagents were compared to those of examples 5-8 using Roche (25 hydroxy vitamin D assay kit, AC-57F 1).
The test result shows that the PBS buffer solution with the concentration of 30mM can provide the optimum buffering environment, and is more favorable for antigen-antibody reaction.
Example 9
A preparation method of a 25-hydroxy vitamin D detection kit comprises a preparation method of a connecting bridge 3, wherein the preparation method of the connecting bridge 3 comprises the following steps: soaking 25 × 30cm cellulose in 40mL of the treatment solution, standing for 24h, drying at 35 deg.C for 12h, and cutting to desired width. The treatment solution comprises 30mM of PBS buffer solution, 0.2% of BSA, 0.7% of PVP, 0.5% of Tween and 0.5% of triton.
The PBS buffer solution has the buffer function of adjusting proper pH, and can fully ensure the biological activity; BSA generally acts as a stabilizer, acting as a "stabilizer" and a "carrier"; PVP, tween 20 and Triton are used as surfactants, so that the PVP, tween 20 and Triton have good non-dissociation property, play a certain role in helping dissolution, can inhibit the automatic polymerization of macromolecules and improve the stability.
Example 10
A preparation method of a 25-hydroxy vitamin D detection kit comprises a preparation method of a connecting bridge 3, wherein the preparation method of the connecting bridge 3 comprises the following steps: soaking 25 × 30cm cellulose in 40mL of the treatment solution, standing for 24h, drying at 35 deg.C for 12h, and cutting to desired width. The treatment solution comprises 30mM of PBS buffer solution, 0.6% of BSA, 0.7% of PVP, 0.5% of Tween and 0.5% of triton.
The PBS buffer solution has the buffer function of adjusting proper pH, and can fully ensure the biological activity; BSA generally acts as a stabilizer, acting as a "stabilizer" and a "carrier"; PVP, tween 20 and Triton are used as surfactants, so that the PVP, tween 20 and Triton have good non-dissociation property, play a certain role in helping dissolution, can inhibit the automatic polymerization of macromolecules and improve the stability.
Example 11
A preparation method of a 25-hydroxy vitamin D detection kit comprises a preparation method of a connecting bridge 3, wherein the preparation method of the connecting bridge 3 comprises the following steps: soaking 25 × 30cm cellulose in 40mL of the treatment solution, standing for 24h, drying at 35 deg.C for 12h, and cutting to desired width. The treatment solution comprises 30mM of PBS buffer solution, 1% of BSA, 0.7% of PVP, 0.5% of Tween and 0.5% of triton.
The PBS buffer solution has the buffer function of adjusting proper pH, and can fully ensure the biological activity; BSA generally acts as a stabilizer, acting as a "stabilizer" and a "carrier"; PVP, tween 20 and Triton are used as surfactants, so that the PVP, tween 20 and Triton are good in non-dissociative property, a certain solubilizing effect is achieved, the autopolymerization of macromolecules can be inhibited, and the stability is improved.
Experiment 2: 20 serum samples were selected and the reagents were compared to those of examples 9-11 using Roche (25 hydroxy vitamin D assay kit, AC-57F 1).
The test results show that the sensitivity is optimal when 0.6% BSA is used in example 10.
Example 12
The preparation method of the 25-hydroxy vitamin D detection kit comprises a preparation method of a connecting bridge 3, wherein the preparation method of the connecting bridge 3 comprises the following steps: soaking 25 × 30cm cellulose in 40mL of the treatment solution, standing for 24h, drying at 35 deg.C for 12h, and cutting to desired width. The treatment solution comprises 30mM of PBS buffer solution, 0.6% of BSA, 0.5% of PVP, 0.5% of Tween and 0.5% of triton.
The PBS buffer solution has the buffer function of adjusting proper pH, and can fully ensure the biological activity; BSA generally acts as a stabilizer, acting as a "stabilizer" and a "carrier"; PVP, tween 20 and Triton are used as surfactants, so that the PVP, tween 20 and Triton are good in non-dissociative property, a certain solubilizing effect is achieved, the autopolymerization of macromolecules can be inhibited, and the stability is improved.
Example 13
The preparation method of the 25-hydroxy vitamin D detection kit comprises a preparation method of a connecting bridge 3, wherein the preparation method of the connecting bridge 3 comprises the following steps: soaking 25 × 30cm cellulose in 40mL of the treatment solution, standing for 24h, drying at 35 deg.C for 12h, and cutting to desired width. The treatment solution comprises 30mM of PBS buffer solution, 0.6% of BSA, 0.8% of PVP, 0.5% of Tween and 0.5% of triton.
The PBS buffer solution has the buffer function of adjusting proper pH, and can fully ensure the biological activity; BSA generally acts as a stabilizer, acting as a "stabilizer" and a "carrier"; PVP, tween 20 and Triton are used as surfactants, so that the PVP, tween 20 and Triton have good non-dissociation property, play a certain role in helping dissolution, can inhibit the automatic polymerization of macromolecules and improve the stability.
Example 14
The preparation method of the 25-hydroxy vitamin D detection kit comprises a preparation method of a connecting bridge 3, wherein the preparation method of the connecting bridge 3 comprises the following steps: soaking 25 × 30cm cellulose in 40mL of the treatment solution, standing for 24h, drying at 35 deg.C for 12h, and cutting to desired width. The treatment solution comprises 30mM of PBS buffer solution, 0.6% of BSA, 1.2% of PVP, 0.5% of Tween and 0.5% of triton.
The PBS buffer solution has the buffer function of adjusting proper pH, and can fully ensure the biological activity; BSA generally acts as a stabilizer, acting as a "stabilizer" and a "carrier"; PVP, tween 20 and Triton are used as surfactants, so that the PVP, tween 20 and Triton have good non-dissociation property, play a certain role in helping dissolution, can inhibit the automatic polymerization of macromolecules and improve the stability.
Experiment 3: 20 serum samples were selected and the reagents were compared to examples 12-14 using Roche (25 hydroxy vitamin D assay kit, AC-57F 1) reagent.
The test results show that the sensitivity is optimal when 0.5% PVP is used in example 13.
Example 15
The preparation method of the 25-hydroxy vitamin D detection kit comprises a preparation method of a connecting bridge 3, wherein the preparation method of the connecting bridge 3 comprises the following steps: soaking 25 × 30cm cellulose in 40mL of the treatment solution, standing for 24h, drying at 35 deg.C for 12h, and cutting to desired width. The treatment solution comprises 30mM of PBS buffer solution, 0.6% of BSA, 0.5% of PVP, 0.2% of Tween and 0.5% of triton.
The PBS buffer solution has the buffer function of adjusting proper pH, and can fully ensure the biological activity; BSA generally acts as a stabilizer, acting as a "stabilizer" and a "carrier"; PVP, tween 20 and Triton are used as surfactants, so that the PVP, tween 20 and Triton have good non-dissociation property, play a certain role in helping dissolution, can inhibit the automatic polymerization of macromolecules and improve the stability.
Example 16
A preparation method of a 25-hydroxy vitamin D detection kit comprises a preparation method of a connecting bridge 3, wherein the preparation method of the connecting bridge 3 comprises the following steps: soaking 25 × 30cm cellulose in 40mL of the treatment solution, standing for 24h, drying at 35 deg.C for 12h, and cutting to desired width. The treatment solution comprises 30mM of PBS buffer solution, 0.6% of BSA, 0.5% of PVP, 0.5% of Tween and 0.5% of triton.
The PBS buffer solution has the buffer function of adjusting proper pH, and can fully ensure the biological activity; BSA generally acts as a stabilizer, acting as a "stabilizer" and a "carrier"; PVP, tween 20 and Triton are used as surfactants, so that the PVP, tween 20 and Triton have good non-dissociation property, play a certain role in helping dissolution, can inhibit the automatic polymerization of macromolecules and improve the stability.
Example 17
The preparation method of the 25-hydroxy vitamin D detection kit comprises a preparation method of a connecting bridge 3, wherein the preparation method of the connecting bridge 3 comprises the following steps: soaking 25 × 30cm cellulose in 40mL of the treatment solution, standing for 24h, drying at 35 deg.C for 12h, and cutting to the desired width. The treatment solution comprises 30mM of PBS buffer solution, 0.6% of BSA, 0.5% of PVP, 20% of Tween and 0.5% of triton.
The PBS buffer solution has the buffer function of adjusting proper pH, and can fully ensure the biological activity; BSA generally acts as a stabilizer, acting as a "stabilizer" and a "carrier"; PVP, tween 20 and Triton are used as surfactants, so that the PVP, tween 20 and Triton have good non-dissociation property, play a certain role in helping dissolution, can inhibit the automatic polymerization of macromolecules and improve the stability.
Experiment 4: 20 serum samples were selected and the reagents were compared to those of examples 15-17 using Roche (25 hydroxy vitamin D assay kit, AC-57F 1).
The test results show that the sensitivity is optimal when 0.5% tween 20 is used in example 16.
Example 18
A preparation method of a 25-hydroxy vitamin D detection kit comprises a preparation method of a connecting bridge 3, wherein the preparation method of the connecting bridge 3 comprises the following steps: soaking 25 × 30cm cellulose in 40mL of the treatment solution, standing for 24h, drying at 35 deg.C for 12h, and cutting to the desired width. The treatment solution comprises 30mM of PBS buffer solution, 0.6% of BSA, 0.5% of PVP, 0.5% of Tween and 0.2% of triton.
The PBS buffer solution has the buffer function of adjusting proper pH, and can fully ensure the biological activity; BSA generally acts as a stabilizer, acting as a "stabilizer" and a "carrier"; PVP, tween 20 and Triton are used as surfactants, so that the PVP, tween 20 and Triton are good in non-dissociative property, a certain solubilizing effect is achieved, the autopolymerization of macromolecules can be inhibited, and the stability is improved.
Example 19
The preparation method of the 25-hydroxy vitamin D detection kit comprises a preparation method of a connecting bridge 3, wherein the preparation method of the connecting bridge 3 comprises the following steps: soaking 25 × 30cm cellulose in 40mL of the treatment solution, standing for 24h, drying at 35 deg.C for 12h, and cutting to desired width. The treatment solution comprises 30mM of PBS buffer solution, 0.6% of BSA, 0.5% of PVP, 0.5% of Tween and 0.8% of triton.
The PBS buffer solution has the buffer function of adjusting proper pH, and can fully ensure the biological activity; BSA generally acts as a stabilizer, acting as a "stabilizer" and a "carrier"; PVP, tween 20 and Triton are used as surfactants, so that the PVP, tween 20 and Triton have good non-dissociation property, play a certain role in helping dissolution, can inhibit the automatic polymerization of macromolecules and improve the stability.
Example 20
A preparation method of a 25-hydroxy vitamin D detection kit comprises a preparation method of a connecting bridge 3, wherein the preparation method of the connecting bridge 3 comprises the following steps: soaking 25 × 30cm cellulose in 40mL of the treatment solution, standing for 24h, drying at 35 deg.C for 12h, and cutting to the desired width. The treatment solution comprises 30mM of PBS buffer solution, 0.6% of BSA, 0.5% of PVP, 0.5% of Tween and 1.5% of triton.
The PBS buffer solution has the buffer function of adjusting proper pH, and can fully ensure the biological activity; BSA generally acts as a stabilizer, acting as a "stabilizer" and a "carrier"; PVP, tween 20 and Triton are used as surfactants, so that the PVP, tween 20 and Triton are good in non-dissociative property, a certain solubilizing effect is achieved, the autopolymerization of macromolecules can be inhibited, and the stability is improved.
Experiment 5: 20 serum samples were selected and the reagents were compared to examples 18-20 using Roche (25 hydroxy vitamin D assay kit, AC-57F 1) reagent.
The test results show that the sensitivity is optimal when 0.8% triton is used in example 19.
The data of experiments 1-5 show that the consistency of the data of examples 5-20 and Roche measurement values is good, and the consistency rate of the invention is higher.
To verify the stability of the 25-hydroxyvitamin D assay kit of the present invention, example 19 was tested for real-time stability, high temperature stability at failure, and transport stability.
Experiment 6: the test strips of example 19 were placed at 2 ℃ for 6, 12, 18, and 24 months, respectively, and 10 serum samples were tested, and the test results were as follows.
The test results show that the test strip in example 19 is placed at 2 ℃ and normal temperature for 6, 12, 18 and 24 months respectively, the detection results are consistent, and the real-time stability of example 19 is determined to be 24 months.
Experiment 7: the test paper strip of example 19 is placed at 45 ℃ for 30d, 60d and 90d, and at 55 ℃ for 30d and 60d, respectively, and compared with the normal temperature, 10 serum samples are tested, and the test results are as follows.
The test result shows that the test strip in example 19 is placed at 45 ℃ for 30d, 60d and 90d, and placed at 55 ℃ for 30d and 60d respectively, the test result is consistent with the test result when the test strip is placed at normal temperature, and the high-temperature damage stability meets the requirement.
Experiment 8: the test paper strip of example 19 was placed at-20 ℃ and 55 ℃ for 7 days and 15 days, respectively, and compared with the normal temperature, 10 serum samples were tested, and the test results are as follows.
The test result shows that the test strip in example 19 is placed for 7d and 15d at-20 ℃ and 55 ℃ respectively, and the test result is consistent with the test result of the test strip placed for 7d and 15d at normal temperature, so that the transportation stability meets the requirement.
The data of experiments 6-8 show that the invention has good stability.
Finally, it should be noted that: the above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and the embodiments are examples, wherein the details that are not described are all the common general knowledge of those skilled in the art, and the technical solutions described in the foregoing embodiments can be modified or some technical features can be equivalently replaced by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A25 hydroxy vitamin D detect reagent box which characterized in that: the test strip comprises a PVC plate, wherein the PVC plate sequentially comprises a water absorption area, a reaction area, a connecting bridge, a release area and a sample adding area from left to right, and the connecting bridge is one of polyurethane, glass fiber, cellulose and a polyester film.
2. The 25 hydroxyvitamin D detection kit according to claim 1, wherein: the connecting bridge is cellulose.
3. The 25 hydroxyvitamin D detection kit according to claim 2, wherein: the preparation method of the connecting bridge comprises the following steps: soaking the cellulose in 40mL of treatment solution, standing for 24h, drying at 35 ℃ for 12h, and cutting to the required width.
4. The 25 hydroxyvitamin D detection kit according to claim 3, wherein: the treatment solution comprises 20-50mM of PBS buffer solution, 0.2-1% of BSA, 0.5-1.2% of PVP, 0.2-1% of Tween 20, and 0.2-1.5% of Triton.
5. The 25 hydroxyvitamin D detection kit according to claim 4, wherein: the treatment solution comprises 30mM of PBS buffer solution, 0.6% of BSA, 0.5% of PVP, 0.5% of Tween and 0.8% of triton.
6. The method for preparing a 25-hydroxy vitamin D detection kit according to claim 1, wherein the method comprises the following steps: the length of the connecting bridge is 6mm.
7. The 25 hydroxyvitamin D detection kit according to claim 6, wherein: one end overlap joint in the reaction zone of water absorption district is served, and the one end overlap joint of connecting the bridge is served in the other end of reaction zone, and the one end overlap joint in the release district is served in the other end of connecting the bridge, and the one end overlap joint in the sample addition district is served in the other end in the release district.
8. The 25 hydroxyvitamin D detection kit according to claim 7, wherein: the lap joint width of the connecting bridge and the reaction zone is 0.7-1mm, and the lap joint width of the release zone and the connecting bridge is 1mm.
9. The 25 hydroxyvitamin D detection kit according to claim 1, wherein: the water absorption area is water absorption paper, the reaction area is a nitrocellulose membrane coated with 25 hydroxy vitamin D antigen, the release area is a gold-labeled solid-phase pad with a solid phase containing 25 hydroxy vitamin D antibody, and the sample adding area is a sample pad.
10. The 25 hydroxyvitamin D detection kit according to claim 1, wherein: the length of the water absorption area is 22mm, the length of the reaction area is 20mm, the length of the release area is 5mm, the length of the sample addition area is 14mm, the overlapping width of the water absorption area and the reaction area is 1mm, and the overlapping width of the sample addition area and the release area is 1.5-2mm.
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Application publication date: 20230328 |