CN109459421B - Serum vitamin A detection kit and detection method thereof - Google Patents
Serum vitamin A detection kit and detection method thereof Download PDFInfo
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- CN109459421B CN109459421B CN201910037689.8A CN201910037689A CN109459421B CN 109459421 B CN109459421 B CN 109459421B CN 201910037689 A CN201910037689 A CN 201910037689A CN 109459421 B CN109459421 B CN 109459421B
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- 238000001514 detection method Methods 0.000 title claims abstract description 62
- 229940045997 vitamin a Drugs 0.000 title claims abstract description 51
- FPIPGXGPPPQFEQ-UHFFFAOYSA-N 13-cis retinol Natural products OCC=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-UHFFFAOYSA-N 0.000 title claims abstract description 49
- FPIPGXGPPPQFEQ-BOOMUCAASA-N Vitamin A Natural products OC/C=C(/C)\C=C\C=C(\C)/C=C/C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-BOOMUCAASA-N 0.000 title claims abstract description 49
- FPIPGXGPPPQFEQ-OVSJKPMPSA-N all-trans-retinol Chemical compound OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-OVSJKPMPSA-N 0.000 title claims abstract description 49
- 235000019155 vitamin A Nutrition 0.000 title claims abstract description 49
- 239000011719 vitamin A Substances 0.000 title claims abstract description 49
- 210000002966 serum Anatomy 0.000 title claims abstract description 32
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 52
- 239000012086 standard solution Substances 0.000 claims abstract description 19
- 239000006228 supernatant Substances 0.000 claims abstract description 16
- 206010070834 Sensitisation Diseases 0.000 claims abstract description 14
- 230000008313 sensitization Effects 0.000 claims abstract description 14
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 11
- 230000005284 excitation Effects 0.000 claims abstract description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 5
- 238000011534 incubation Methods 0.000 claims abstract description 4
- 238000000034 method Methods 0.000 claims description 17
- 238000004806 packaging method and process Methods 0.000 claims description 11
- 230000008093 supporting effect Effects 0.000 claims description 10
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical group CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 8
- 238000005192 partition Methods 0.000 claims description 6
- 238000012856 packing Methods 0.000 claims description 5
- 239000004094 surface-active agent Substances 0.000 claims description 5
- 238000012360 testing method Methods 0.000 claims description 5
- 239000003945 anionic surfactant Substances 0.000 claims description 4
- 239000004064 cosurfactant Substances 0.000 claims description 4
- 239000000243 solution Substances 0.000 claims description 4
- 239000003093 cationic surfactant Substances 0.000 claims description 3
- 238000002474 experimental method Methods 0.000 claims description 3
- 239000000693 micelle Substances 0.000 claims description 2
- 239000004530 micro-emulsion Substances 0.000 claims description 2
- 239000002736 nonionic surfactant Substances 0.000 claims description 2
- 210000000352 storage cell Anatomy 0.000 claims 1
- 239000011148 porous material Substances 0.000 abstract description 3
- 239000000123 paper Substances 0.000 description 9
- 239000006260 foam Substances 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 239000011111 cardboard Substances 0.000 description 3
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 3
- 229940088594 vitamin Drugs 0.000 description 3
- 229930003231 vitamin Natural products 0.000 description 3
- 235000013343 vitamin Nutrition 0.000 description 3
- 239000011782 vitamin Substances 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 230000007812 deficiency Effects 0.000 description 2
- 230000006866 deterioration Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000002848 electrochemical method Methods 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 150000003722 vitamin derivatives Chemical class 0.000 description 2
- 208000032170 Congenital Abnormalities Diseases 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 206010058314 Dysplasia Diseases 0.000 description 1
- 206010060919 Foetal malformation Diseases 0.000 description 1
- 208000020241 Neonatal disease Diseases 0.000 description 1
- 208000037129 Newborn Diseases Infant Diseases 0.000 description 1
- 206010057190 Respiratory tract infections Diseases 0.000 description 1
- 208000007502 anemia Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000013016 damping Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002795 fluorescence method Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 239000011785 micronutrient Substances 0.000 description 1
- 235000013369 micronutrients Nutrition 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 230000002028 premature Effects 0.000 description 1
- 238000002791 soaking Methods 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
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N21/643—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" non-biological material
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- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Optics & Photonics (AREA)
- Molecular Biology (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a serum vitamin A detection kit and a detection method thereof, comprising the following steps: (1) Taking 20 mu l of vitamin A standard solution with the concentration of 10 mu g/dl, 20 mu g/dl and 30 mu g/dl and serum samples to be tested, respectively marking the standard group A, the standard group B, the standard group C and the experimental group, adding 20 mu l of deproteinizing agent into each, centrifuging for 15min at 3000r/min, and taking supernatant; (2) Sequentially adding the supernatant into a 96-hole micro-pore plate, respectively adding 180 mu l of sensitization reagent, and incubating for 3 hours in a water bath kettle at 60 ℃; (3) After the incubation is finished, measuring fluorescence values with excitation wavelength of 345nm and emission wavelength of 480nm by using a microplate fluorescence analyzer, fitting a standard curve by using the concentration and the fluorescence values of each gradient of the standard solution of vitamin A, and substituting the fluorescence values of the serum sample to be detected into the standard curve to calculate the content of the vitamin A. The sensitization reagent adopted by the invention is used for carrying out fluorescence sensitization, thus realizing the quantitative detection of the vitamin A in a trace serum sample (20 mu l).
Description
Technical Field
The invention relates to the technical field of kit detection, in particular to a serum vitamin A detection kit and a detection method thereof.
Background
Vitamin A (VA) is one of important vitamins required by human bodies, is an indispensable micronutrient in the growth and development process of children, and the VA deficiency can have great influence on the bodies of the children: can cause fetal malformation, premature infant dysplasia, neonatal disease increase, influence the growth and development of children, cause anemia, immune function decline, respiratory tract infection, diarrhea incidence and disease death rate rise, and has obvious influence on young children. Thus, knowing the harm of VA deficiency, and early intervention and treatment of VA deficient people is an important factor for ensuring healthy growth of children.
VA is very easily oxidized in air, so that the detection quality can be ensured only by realizing rapid detection. Currently available VA detection methods include a fluorescence method, a high performance liquid chromatography method and an electrochemical method. The methods have common application defects such as large required sample size, easy influence on detection results caused by oxidative deterioration after sample collection, incapability of rapid batch detection and the like. Therefore, the method is not suitable for vitamin content detection of infants and children. The vitamin detection for adults has no problem of sample size, but also has the problem of easy deterioration after sample collection.
As is well known, currently available methods for detecting vitamin A include fluorescence, HPLC and electrochemical methods. These methods generally require the use of a detection kit, which is present in large quantities in order to meet the requirements of the detection, however, the known kits generally have several drawbacks:
(1) The internal pattern of the detection kit is unreasonable, and the reagent and the instrument are usually arranged on the same layer, so that the reagent and the instrument are placed in disorder, and a detector cannot take the reagent and the instrument rapidly during working, thereby increasing the workload of the detector and reducing the detection speed;
(2) The detection kit is softer in texture, and is easy to squeeze in the transportation process, so that the reagent and the instrument are subjected to position deviation, and even the reagent instrument is damaged due to mutual collision;
(3) The detection plate and the reagent are unreasonably placed in the detection process of the detection personnel, so that the detection efficiency is greatly affected, and the detection efficiency is reduced.
Disclosure of Invention
The invention aims to provide a serum vitamin A detection kit for solving the problems in the background technology.
In order to achieve the above purpose, the present invention provides the following technical solutions: the utility model provides a serum vitamin A detect reagent box, includes stereoplasm support body, packing carton and pick-up plate, the stereoplasm support body is rectangular frame structure to the inside of stereoplasm support body has first storing chamber, second storing chamber and third storing chamber through the partition frame that evenly sets up, and the outside of stereoplasm support body evenly wraps up simultaneously has the packing carton, the inside movable mounting in third storing chamber has first drawer, and the both sides in second storing chamber are provided with second drawer and third drawer respectively, and first traction area is all installed to the centre of first drawer, second drawer and third drawer one end to the inside of second drawer is provided with the reagent board, and the inside of third drawer is provided with the storage frame, evenly is provided with the fixed orifices on the storage frame simultaneously, the centre at reagent board top is equipped with the recess, and the inside packing of recess has foam to evenly set up first reagent hole and second reagent hole, and the both ends at reagent board top all are fixed with the roof frame, the inside in first storing chamber is provided with the second drawer and the inside of card board, and the inside of the same side that is provided with the folding cover plate, the inside of the card board and the same side that is provided with the inside of folding cover plate, the inside of both sides that the card board and the same side cover plate are provided with the equal to the inside of folding cover plate, the inside of the cover plate is provided with the same side as the cover plate, and the inside cover plate is provided with the folding cover plate.
Further, a first opening matched with the first drawer is formed in the bottom of one side of the packaging paper box, and a second opening and a third opening matched with the second drawer and the third drawer are formed in the middle of two ends of the packaging paper box.
Further, a second traction belt is arranged in the middle of the two ends of the top of the detection plate.
Further, the number of micropores at the top of the support plate is 96 holes.
Further, storage grids are uniformly arranged in the first drawer through the cross partition plates.
A method for serum VA detection comprising the steps of:
step one: taking 20 mu l of vitamin A standard solution with the concentration of 10 mu g/dl, 20 mu g/dl and 30 mu g/dl and serum samples to be tested, respectively marking the standard group A, the standard group B, the standard group C and the experimental group, carrying out 3 parallel experiments on the standard group A, the standard group B, the standard group C and the experimental group, adding 20 mu l of deproteinizing agent into the standard group A, the standard group B, the standard group C and the experimental group, centrifuging for 15min at 3000r/min, and taking supernatant;
step two: sequentially adding the supernatant in the first step into a 96-hole micro-pore plate, and respectively adding 180 mu l of sensitization reagent, and incubating in a water bath kettle at 60 ℃ for 3 hours;
step three: after the incubation is finished, measuring fluorescence values with excitation wavelength of 345nm and emission wavelength of 480nm by using a microplate fluorescence analyzer, fitting a standard curve by using the concentration and the fluorescence values of each gradient of the standard solution of vitamin A, and substituting the fluorescence values of the serum sample to be detected into the standard curve to calculate the content of the vitamin A.
Further, the standard solution concentration of vitamin A is 10. Mu.g/dl, 20. Mu.g/dl and 30. Mu.g/dl.
Further, the sensitization agent is micelle solution or microemulsion composed of a surfactant and a cosurfactant.
Further, the surfactant is an anionic surfactant, a cationic surfactant or a nonionic surfactant.
Further, the cosurfactant is ethanol or glycol.
Compared with the prior art, the invention has the beneficial effects that:
(1) According to the invention, the hard frame body with the rectangular frame structure is matched with the packaging paper boxes uniformly wrapped on the outer side of the hard frame body to form a complete kit structure, so that the structural strength of the kit can be greatly enhanced, and the condition that the kit is easy to squeeze in the transportation process can be effectively avoided.
(2) According to the invention, the first storage cavity, the second storage cavity and the third storage cavity are uniformly arranged in the partition frame utilized in the hard frame body, the first drawer is movably arranged in the third storage cavity, the second drawer and the third drawer are respectively arranged at two sides of the second storage cavity, so that instruments and reagents can be effectively placed in the drawers of different storage cavities respectively, the placement positions of the instruments and the reagents are effectively distinguished, the pattern is clear, the level is clear, and the placement of articles is ordered.
(3) According to the invention, the detection plate and the reagent plate which are mutually combined are arranged, so that the detection plate can be arranged at the top of the reagent plate in the detection process, the use of detection personnel is facilitated, and the improvement of the detection speed is facilitated.
(4) The invention adopts the supporting plate formed by 96 holes and micropores to detect the vitamin A content, realizes rapid detection, and has the detection efficiency greatly higher than that of the traditional detection method.
(5) The sensitization reagent adopted by the invention is used for carrying out fluorescence sensitization, thus realizing the quantitative detection of the vitamin A in a trace serum sample (20 mu l).
(6) The invention adopts the sensitization reagent with strong stability, can stabilize fluorescence for 2 hours, and ensures the detection stability by using the sensitization reagent for fluorescence sensitization.
Drawings
FIG. 1 is a schematic diagram of the overall structure of the present invention;
FIG. 2 is a schematic view of a rigid frame structure according to the present invention;
FIG. 3 is a schematic diagram showing the combined use structure of the detection plate and the reagent plate according to the present invention.
In the figure: 1-a first drawer; 2-reagent plate; 3-a second drawer; 4-micropores; 5-detecting plate; 6-an elastic belt; 7-an upper cover plate; 8-clamping holes; 9-packaging paper boxes; 10-a third drawer; 11-a storage rack; 12-fixing holes; 13-a first traction belt; 14-a second traction belt; 15-top rack; 16-a support plate; 17-supporting frames; 18-clamping grooves; 19-grooves; 20-a first reagent well; 21-soaking cotton; 22-a second reagent well; 23-a first storage cavity; 24-a third storage cavity; 25-a hard frame body; 26-a second storage cavity; 27-spacer.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Referring to fig. 1-3, the present invention provides a technical solution: a serum vitamin A detection kit comprises a hard frame body 25, a packaging paper box 9 and a detection plate 5, wherein the hard frame body 25 is of a rectangular frame structure, a first storage cavity 23, a second storage cavity 26 and a third storage cavity 24 are formed in the hard frame body 25 through a partition frame 27 which is uniformly arranged, the packaging paper box 9 is uniformly wrapped on the outer side of the hard frame body 25, a first drawer 1 is movably arranged in the third storage cavity 24, storage grids are uniformly arranged in the first drawer 1 through cross partition plates, objects such as rubber head droppers and test tubes are stored in the storage grids, a second drawer 3 and a third drawer 10 are respectively arranged on two sides of the second storage cavity 26, a first opening which is mutually matched with the first drawer 1 is formed in the bottom of one side of the packaging paper box 9, a second opening and a third opening which are mutually matched with the second drawer 3 and the third drawer 10 are formed in the middle of two ends of the packaging paper box 9, the first traction belt 13 is arranged in the middle of one end of the first drawer 1, the second drawer 3 and the third drawer 10, the reagent plate 2 is arranged in the second drawer 3, the storage rack 11 is arranged in the third drawer 10, the fixing holes 12 are uniformly arranged on the storage rack 11, the fixing holes 12 are used for fixing medicaments, the middle of the top of the reagent plate 2 is provided with a groove 19, foam 21 is filled in the groove 19, the foam 21 has good damping function and avoids reagent bottle damage, the first reagent hole 20 and the second reagent hole 22 are uniformly arranged in the foam 21, the reagent bottle filled with vitamin A standard solution, sensitization reagent and deproteinizing agent is arranged in the first reagent hole 20 or the second reagent hole 22, the top rack 15 is fixed at both ends of the top of the reagent plate 2, the detection plate 5 is arranged in the first storage cavity 23, the second traction belt 14 is installed in the middle of the two ends of the top of the detection plate 5, the detection plate 5 is conveniently taken out of the first storage cavity 23 through the arrangement of the second traction belt 14, clamping grooves 18 matched with the top frame 15 are formed in the two ends of the bottom of the detection plate 5, a supporting frame 17 is fixed to the bottom of the detection plate 5 on the inner side of the clamping grooves 18, in the using process, the detection plate 5 is placed on the top of the reagent plate 2, the clamping grooves 18 on the bottom of the detection plate 5 are sleeved on the top of the top frame 15, the supporting frame 17 is matched, the dual supporting effect is achieved, the combined use stability of the detection plate 5 and the reagent plate 2 is guaranteed, meanwhile, a supporting plate 16 is arranged in the middle of the detection plate 5, the detection plate 5 and the supporting plate 16 are of an integral structure, micropores 4 are uniformly formed in the top of the supporting plate 16, the micropores 4 on the top of the supporting plate 16 are 96 holes, an upper cover plate 7 folded inwards is arranged on the top of one side of the packaging paper box 9, an elastic belt 6 is arranged on the inner side of the upper cover plate 7, a clamping hole 8 is uniformly formed in the joint of the elastic belt 6 and the upper cover plate 7, and the elastic belt 6 is provided with a certain elastic space matched with the clamping hole 8 for clamping test tubes.
Working principle: the rubber head dropper, test tube and other instruments are placed in the storage grid in the first drawer 1, the reagent bottles containing vitamin A standard solution, sensitization reagent and deproteinizing agent with the concentration of 10 mug/dl, 20 mug/dl and 30 mug/dl are placed in the first reagent hole 20 and the second reagent hole 22 in the second drawer 3, in the using process, the reagent plate 2 in the second drawer 3 can be taken out, the second traction belt 14 is utilized to take out the detection plate 5 from the first storage cavity 23, and the detection plate 5 is placed on the top of the reagent plate 2, at the moment, the clamping groove 18 at the bottom of the detection plate 5 is sleeved on the top of the top frame 15, so that the combination use of the detection plate 5 and the reagent plate 2 is realized, the reagent can be conveniently taken out by a detector, and the improvement of the detection speed is facilitated.
A method for serum VA detection comprising the steps of:
step one: taking 20 mu l of vitamin A standard solution with the concentration of 10 mu g/dl, 20 mu g/dl and 30 mu g/dl and serum samples to be tested, respectively marking the standard group A, the standard group B, the standard group C and the experimental group, carrying out 3 parallel experiments on the standard group A, the standard group B, the standard group C and the experimental group, adding 20 mu l of deproteinizing agent into the standard group A, the standard group B, the standard group C and the experimental group, centrifuging for 15min at 3000r/min, and taking supernatant;
step two: sequentially adding the supernatant in the first step into a 96-hole micro-pore plate, and respectively adding 180 mu l of sensitization reagent, and incubating in a water bath kettle at 60 ℃ for 3 hours;
step three: after the incubation is finished, measuring fluorescence values with excitation wavelength of 345nm and emission wavelength of 480nm by using a microplate fluorescence analyzer, fitting a standard curve by using the concentration and the fluorescence values of each gradient of the standard solution of vitamin A, and substituting the fluorescence values of the serum sample to be detected into the standard curve to calculate the content of the vitamin A.
Example 1:
step one: taking 20 mu l of vitamin A standard solution (10 mu g/dl, 20 mu g/dl and 30 mu g/dl) and serum samples to be tested respectively, adding deproteinizing agents respectively, carrying out deproteinizing treatment, centrifuging at a volume ratio of 1:1, 3000r/min for 15min, and taking supernatant.
Step two: the supernatant was added sequentially to 96-well microwell plates and 180. Mu.l of a sensitizer (anionic surfactant and ethanol mixture) was added, respectively, and incubated in a 60℃water bath for 3h.
Step three: detecting by using a microplate fluorescence analyzer, measuring the fluorescence value of the sample with excitation wavelength of 345nm and emission wavelength of 480nm, fitting a standard curve by the concentration and the fluorescence value of each gradient of the vitamin A standard solution, and substituting the fluorescence value of the serum sample to be detected into the standard curve to calculate the vitamin A content.
Example 2:
step one: taking 20 mu l of vitamin A standard solution (10 mu g/dl, 20 mu g/dl and 30 mu g/dl) and serum samples to be tested respectively, adding deproteinizing agents respectively, carrying out deproteinizing treatment, centrifuging at a volume ratio of 1:1, 3000r/min for 15min, and taking supernatant.
Step two: the supernatant was added sequentially to 96-well microwell plates and 180. Mu.l of a sensitizer (cationic surfactant and ethanol mixture) was added, respectively, and incubated in a 60℃water bath for 3h.
Step three: detecting by using a microplate fluorescence analyzer, measuring the fluorescence value of the sample with excitation wavelength of 345nm and emission wavelength of 480nm, fitting a standard curve by the concentration and the fluorescence value of each gradient of the vitamin A standard solution, and substituting the fluorescence value of the serum sample to be detected into the standard curve to calculate the vitamin A content.
Example 3:
step one: taking 20 mu l of vitamin A standard solution (10 mu g/dl, 20 mu g/dl and 30 mu g/dl) and serum samples to be tested respectively, adding deproteinizing agents respectively, carrying out deproteinizing treatment, centrifuging at a volume ratio of 1:1, 3000r/min for 15min, and taking supernatant.
Step two: the supernatants were added sequentially to 96-well microplates and 180 μl of sensitizer (non-surfactant and glycol mixture) were added, respectively, and incubated for 3h in a 60℃water bath.
Step three: detecting by using a microplate fluorescence analyzer, measuring the fluorescence value of the sample with excitation wavelength of 345nm and emission wavelength of 480nm, fitting a standard curve by the concentration and the fluorescence value of each gradient of the vitamin A standard solution, and substituting the fluorescence value of the serum sample to be detected into the standard curve to calculate the vitamin A content.
Example 4:
step one: taking 20 mu l of vitamin A standard solution (10 mu g/dl, 20 mu g/dl and 30 mu g/dl) and serum samples to be tested respectively, adding deproteinizing agents respectively, carrying out deproteinizing treatment, centrifuging at a volume ratio of 1:1, 3000r/min for 15min, and taking supernatant.
Step two: the supernatants were added sequentially to 96-well microplates and 180 μl of sensitizer (anionic surfactant and glycol mixture) were added, respectively, and incubated in a 60 ℃ water bath for 3h.
Step three: detecting by using a microplate fluorescence analyzer, measuring the fluorescence value of the sample with excitation wavelength of 345nm and emission wavelength of 480nm, fitting a standard curve by the concentration and the fluorescence value of each gradient of the vitamin A standard solution, and substituting the fluorescence value of the serum sample to be detected into the standard curve to calculate the vitamin A content.
In summary, 10 repeated analyses were performed on the same serum sample, the inter-batch variation coefficient was 4.97%, and 10 analyses were performed on the same serum sample in parallel, and the intra-batch variation coefficient was 2.87%.
The standard curve has good linearity when the vitamin A concentration is 1-50 mug/dl.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.
Claims (7)
1. The utility model provides a serum vitamin A detection kit, its characterized in that, includes stereoplasm support body (25), stereoplasm support body (25) are rectangular frame structure, just the inside of stereoplasm support body (25) is equipped with first storing chamber (23), second storing chamber (26) and third storing chamber (24) through the frame (27) that evenly sets up, the outside of stereoplasm support body (25) is evenly wrapped up there is packing carton (9), the inside movable mounting of third storing chamber (24) has first drawer (1), the both sides of second storing chamber (26) are provided with second drawer (3) and third drawer (10) respectively, first drawer (13) are all installed in the centre of first drawer (1), second drawer (3) and third drawer (10) one end, just the inside of second drawer (3) is provided with reagent board (2), the inside of third drawer (10) is provided with frame (11), simultaneously on storing frame (11) evenly have reagent board (12), reagent bubble (21) are equipped with in the top (21) is equipped with in the centre of a hole (21) and two tops (19) are equipped with, reagent bubble (21) are all equipped with in the top (19), the inside of first storing chamber (23) is provided with pick-up plate (5), both ends of pick-up plate (5) bottom all are provided with draw-in groove (18) that match each other with roof-rack (15), pick-up plate (5) bottom fixedly connected with support frame (17) of draw-in groove (18) inboard, the centre of pick-up plate (5) is provided with backup pad (16), the top of backup pad (16) evenly is provided with micropore (4), the top of packing carton (9) one side is equipped with upper cover plate (7) of inboard folding, be provided with elastic webbing (6) on the inside wall of upper cover plate (7), the junction of elastic webbing (6) and upper cover plate (7) evenly is provided with clip hole (8);
the bottom of one side of the packaging paper box (9) is provided with a first opening matched with the first drawer (1), and the middle parts of two ends of the packaging paper box (9) are provided with a second opening and a third opening matched with the second drawer (3) and the third drawer (10).
2. The serum vitamin a test kit according to claim 1, wherein the second traction belt (14) is mounted in the middle of both ends of the top of the test plate (5).
3. The serum vitamin a detection kit according to claim 1, wherein the first drawer (1) is internally and uniformly provided with storage cells through cross-shaped partition plates.
4. A detection method using the kit according to claims 1 to 3, characterized by comprising the steps of:
step one: taking 20 mu l of vitamin A standard solution with the concentration of 10 mu g/dl, 20 mu g/dl and 30 mu g/dl and serum samples to be tested, respectively marking the standard group A, the standard group B, the standard group C and the experimental group, carrying out 3 parallel experiments on the standard group A, the standard group B, the standard group C and the experimental group, adding 20 mu l of deproteinizing agent into the standard group A, the standard group B, the standard group C and the experimental group, centrifuging for 15min at 3000r/min, and taking supernatant;
step two: sequentially adding the supernatant in the first step into a supporting plate with 96 micropores at the top, and respectively adding 180 μl of a sensitization reagent, and incubating in a water bath kettle at 60 ℃ for 3 hours;
step three: after the incubation is finished, a supporting plate fluorescence analyzer with 96 micropores at the top is used for measuring fluorescence values at the excitation wavelength of 345nm and the emission wavelength of 480nm, a standard curve is fitted by the concentration and the fluorescence values of each gradient of the standard solution of vitamin A, and the fluorescence values of the serum sample to be measured are substituted into the standard curve to calculate the content of the vitamin A.
5. The method according to claim 4, wherein the sensitizer is a micelle solution or microemulsion composed of a surfactant and a cosurfactant.
6. The method according to claim 5, wherein the surfactant is an anionic surfactant, a cationic surfactant or a nonionic surfactant.
7. The method of claim 5, wherein the cosurfactant is ethanol or ethylene glycol.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910037689.8A CN109459421B (en) | 2019-01-16 | 2019-01-16 | Serum vitamin A detection kit and detection method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910037689.8A CN109459421B (en) | 2019-01-16 | 2019-01-16 | Serum vitamin A detection kit and detection method thereof |
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6271039B1 (en) * | 1997-12-24 | 2001-08-07 | Kalibrant Limited | Composition for use in fluorescence assay systems |
| CN1456894A (en) * | 2003-06-09 | 2003-11-19 | 江南大学 | Diethylstilbestrol enzyme linked immunoassay reagent box and determination |
| CN1549921A (en) * | 2001-06-29 | 2004-11-24 | ÷ | Assay plates, reader systems and methods for luminescence test measurements |
| AU2011265549A1 (en) * | 2005-12-21 | 2012-02-02 | Meso Scale Technologies, Llc | Assay Modules Having Assay Reagents and Methods of Making and Using Same |
| CN205499819U (en) * | 2016-03-12 | 2016-08-24 | 北京易科拜德科技有限公司 | Reagent kit |
| CN109061181A (en) * | 2018-08-02 | 2018-12-21 | 宁波奥丞生物科技有限公司 | A kind of vascular endothelial growth factor chemiluminescence detection kit |
| CN209559763U (en) * | 2019-01-16 | 2019-10-29 | 岳昆 | A kind of serium vitamin A detection kit |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US10730044B2 (en) * | 2015-10-01 | 2020-08-04 | The Regents Of The University Of Michigan | Assay plate and uses thereof |
-
2019
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Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6271039B1 (en) * | 1997-12-24 | 2001-08-07 | Kalibrant Limited | Composition for use in fluorescence assay systems |
| CN1549921A (en) * | 2001-06-29 | 2004-11-24 | ÷ | Assay plates, reader systems and methods for luminescence test measurements |
| CN1456894A (en) * | 2003-06-09 | 2003-11-19 | 江南大学 | Diethylstilbestrol enzyme linked immunoassay reagent box and determination |
| AU2011265549A1 (en) * | 2005-12-21 | 2012-02-02 | Meso Scale Technologies, Llc | Assay Modules Having Assay Reagents and Methods of Making and Using Same |
| CN205499819U (en) * | 2016-03-12 | 2016-08-24 | 北京易科拜德科技有限公司 | Reagent kit |
| CN109061181A (en) * | 2018-08-02 | 2018-12-21 | 宁波奥丞生物科技有限公司 | A kind of vascular endothelial growth factor chemiluminescence detection kit |
| CN209559763U (en) * | 2019-01-16 | 2019-10-29 | 岳昆 | A kind of serium vitamin A detection kit |
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