CN201408194Y - ELISA detection kit for trace albumin in human body - Google Patents
ELISA detection kit for trace albumin in human body Download PDFInfo
- Publication number
- CN201408194Y CN201408194Y CN2009201520440U CN200920152044U CN201408194Y CN 201408194 Y CN201408194 Y CN 201408194Y CN 2009201520440 U CN2009201520440 U CN 2009201520440U CN 200920152044 U CN200920152044 U CN 200920152044U CN 201408194 Y CN201408194 Y CN 201408194Y
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- Prior art keywords
- albumin
- phase carrier
- detection kit
- solid phase
- elisa
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- 238000002965 ELISA Methods 0.000 title claims abstract description 29
- 108010088751 Albumins Proteins 0.000 title claims abstract description 22
- 102000009027 Albumins Human genes 0.000 title claims abstract description 22
- 238000001514 detection method Methods 0.000 title claims abstract description 18
- 239000007790 solid phase Substances 0.000 claims abstract description 18
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims abstract description 4
- 229940098773 bovine serum albumin Drugs 0.000 claims abstract description 4
- 239000004793 Polystyrene Substances 0.000 claims description 9
- 108091006905 Human Serum Albumin Proteins 0.000 claims description 6
- 102000008100 Human Serum Albumin Human genes 0.000 claims description 6
- 239000003795 chemical substances by application Substances 0.000 claims description 6
- 229920002223 polystyrene Polymers 0.000 claims description 5
- 210000002700 urine Anatomy 0.000 abstract description 9
- 230000035945 sensitivity Effects 0.000 abstract description 5
- 108090000790 Enzymes Proteins 0.000 abstract description 4
- 102000004190 Enzymes Human genes 0.000 abstract description 4
- 210000002966 serum Anatomy 0.000 abstract description 3
- 238000011084 recovery Methods 0.000 abstract description 2
- 238000000034 method Methods 0.000 description 17
- FRPHFZCDPYBUAU-UHFFFAOYSA-N Bromocresolgreen Chemical compound CC1=C(Br)C(O)=C(Br)C=C1C1(C=2C(=C(Br)C(O)=C(Br)C=2)C)C2=CC=CC=C2S(=O)(=O)O1 FRPHFZCDPYBUAU-UHFFFAOYSA-N 0.000 description 4
- ABIUHPWEYMSGSR-UHFFFAOYSA-N bromocresol purple Chemical compound BrC1=C(O)C(C)=CC(C2(C3=CC=CC=C3S(=O)(=O)O2)C=2C=C(Br)C(O)=C(C)C=2)=C1 ABIUHPWEYMSGSR-UHFFFAOYSA-N 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- UAIUNKRWKOVEES-UHFFFAOYSA-N 3,3',5,5'-tetramethylbenzidine Chemical compound CC1=C(N)C(C)=CC(C=2C=C(C)C(N)=C(C)C=2)=C1 UAIUNKRWKOVEES-UHFFFAOYSA-N 0.000 description 1
- 108090001008 Avidin Proteins 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 230000006287 biotinylation Effects 0.000 description 1
- 238000007413 biotinylation Methods 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000016097 disease of metabolism Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
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- 208000014674 injury Diseases 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 208000017169 kidney disease Diseases 0.000 description 1
- 238000003771 laboratory diagnosis Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
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- Investigating Or Analysing Biological Materials (AREA)
Abstract
The utility model provides an ELISA (enzyme linked immunosorbent assay) detection kit for trace albumin in human body. The detection kit comprises a kit cover, a kit body and an enzyme label plate, wherein the enzyme label plate further includes a solid-phase carrier and an enveloping layer. The solid-phase carrier is a micro-porous plate with a plurality of micropores, and the enveloping layer isattached to the micro-porous surface of the solid-phase carrier and formed by applying mouse anti-human albumin and bovine serum albumin on the micropores. When being used for detecting serum specimens and urine specimens of the human, the detection kit greatly improves the sensitivity; meanwhile, the recovery rate is high, so that the repeatability is good; and the cost is low.
Description
Technical field
The utility model relates to diagnostic reagent application apparatus field, relates in particular to a kind of kit that is used to detect human albumin.
Background technology
Human serum albumins is an important indicator of weighing liver function and body nutrition condition.At present, be used to detect albuminous method and mainly contain bromcresol purple (BCP) method, protein electrophoresis and bromcresol green (BCG) method, wherein BCP and BCG method need be utilized automatic clinical chemistry analyzer, it is easy and simple to handle, quick, but automatic clinical chemistry analyzer costs an arm and a leg, one-time investment is big, and sensitivity is not high, and is not high for the albuminous accuracy in detection of denier; The protein electrophoresis complex operation, and on the low side because of albumin and Ponceau S dyestuff adhesion, often cause the result on the low side, seldom be used at present.Therefore, be necessary to select for use the high method of a kind of economy, sensitivity and accuracy to carry out albumin, the particularly albuminous detection of denier.
Enzyme-linked immunosorbent assay (enzyme linked immunosorbent assay, ELISA) be the laboratory diagnosis method of a kind of susceptibility height of organically combining of the efficient catalytic action principle with the specificity of antigen, antibody mediated immunity reaction and enzyme, high specificity, good reproducibility, can detect the specific proteins of trace in the body fluid sensitively.
The utility model content
The technical problems to be solved in the utility model provides a kind of people's denier albumin ELISA detection kit, and it is easy and simple to handle, and cost is low, and good stability is suitable for albumin detection in blood and the urine, and particularly the denier albumin detects in the urine.
For solving the problems of the technologies described above, the technical solution of the utility model is: a kind of kit that is used for ELISA method detection human albumin, comprise lid, box body and ELISA Plate, wherein ELISA Plate comprises solid phase carrier and bag tegillum again, solid phase carrier is the microwell plate that is provided with a plurality of micropores, the bag tegillum is attached to the micropore surface of solid phase carrier, and the bag tegillum is coated on the micropore successively by mouse anti human albumin monoclonal antibody and bovine serum albumin(BSA) and forms.
Preferably, described solid phase carrier is 96 hole polystyrene agent plate (PS).
Preferably, described solid phase carrier is 48 hole polystyrene agent plate (PS).
Our experiments show that, use the utility model its lowest detection when detecting human serum and urine specimen to be limited to 0.15ng/ml, and traditional full-automatic biochemical BCP or BCG rule are 1ug/ml, have greatly improved sensitivity; Its recovery is 87 ± 15% simultaneously, and repeatability is also good, therefore is specially adapted to the urine microalbumin and detects, and has important clinic value for the generation of liver, injury of kidney and metabolic disease, the monitoring of development; In addition, the utility model need not invested large-scale automatic biochemistry analyzer, and method of operating is simple, and is economical, quick, cost is low.
Description of drawings
Fig. 1 is the utility model kit structural representation;
Fig. 2 is the utility model ELISA Plate structural representation;
Fig. 3 is the utility model ELISA Plate cut-open view.
Embodiment
As shown in Figure 1, the utility model kit is by lid 4, and ELISA Plate 5 and box body 6 are formed.
As shown in Figures 2 and 3, ELISA Plate 5 is made up of solid phase carrier 1 and bag tegillum 3, and micropore 2 is arranged on the solid phase carrier 1, and bag tegillum 3 is attached on the micropore 2.
Preparation method of the present utility model can be: adopt 96 hole polystyrene agent plate (PS) as solid phase carrier, be placed under the ultraviolet light irradiation earlier 2 hours, make the activation of PS plate.Bag is by certain density mouse anti human albumin monoclonal antibody earlier on the agent plate capillary strip, and 4 ℃ are spent the night, and makes bag by good ELISA Plate through washing after the sealing of plate and bovine serum albumin(BSA) is handled, and detects in order to sample.
Using method of the present utility model is: successively with the anti-human albumin polyclonal antibody of biotinylation rabbit, Avidin horseradish peroxidase (HRP) is as detectable A and B, with 3,3 ', 5,5 '-tetramethyl benzidine (TMB) and sulfuric acid is respectively as substrate and stop buffer, finishes to be used for the use that the ELISA method detects the kit of human albumin.
Embodiment 1: denier albumin ELISA detection kit and large-scale automatic biochemistry analyzer detection by quantitative people's definite value (35.5ng/ml) urine specimen respectively of should choosing.
1, people's denier albumin ELISA detectable cassette method operation: 1. the kit balance is to room temperature.2. application of sample: establish blank well, gauge orifice, testing sample hole respectively.Blank well adds sample dilution 100ul, and surplus hole adds standard items (Alb) or definite value sample 100ul respectively, and overlay film was placed 120 minutes for 37 ℃.Standard items concentration is respectively: 40ng/ml, 20ng/ml, 10ng/ml, 5ng/ml, 2.5ng/ml, 1.25ng/ml, 0.625ng/ml.3. discard liquid, dry, every hole adds detectable A working fluid 100ul, and overlay film was placed 60 minutes for 37 ℃.4. discard liquid in the hole, wash plate 3 times, dry incubation 60 minutes.5. every hole adds detects liquid reagent B working fluid 100ul, and overlay film was placed 60 minutes for 37 ℃.6. discard liquid in the hole, wash plate 5 times, dry.7. every hole adds TMB solution 90ul, and 37 ℃ of lucifuges developed the color 30 minutes.8. every in regular turn hole adds stop bath 50ul, with the optical density (OD value) of enzyme connection instrument in each hole of 450nm wavelength measurement.
2, large-scale automatic biochemistry analyzer detects same above-mentioned urine specimen
According to the instrument requirement, last sample, the operation instrument, instrument is got the result automatically.
3, result: see Table 1
Table 1. different experiments method testing result relatively
Embodiment 2: denier albumin ELISA detection kit and large-scale automatic biochemistry analyzer detection by quantitative people's definite value (4.86ug/ml) urine specimen respectively of should choosing.
Method and operation be with embodiment 1, but need carry out 200 times of dilutions to sample when detecting with the ELISA method.
Result: see Table 2
Use the utility model ELISA method as can be seen from the data of table 1 and table 2 and detect albumin content the urine, its accuracy height.When albumin content is higher in the sample and large-scale automatic biochemistry analyzer relatively do not have significant difference, but albumin content is atomic hour in sample, the biochemical instruments method can't be measured, but the used ELISA method of the utility model but can detect the albumin of denier, greatly improved the sensitivity that albumin detects, monitoring for generation, development and the medication of prompting relevant disease has the important clinical meaning.
Claims (3)
1. people's denier albumin ELISA detection kit, comprise lid, box body and ELISA Plate, wherein ELISA Plate comprises solid phase carrier and bag tegillum again, it is characterized in that: described solid phase carrier is the microwell plate that is provided with a plurality of micropores, described bag tegillum is attached to the micropore surface of described solid phase carrier, and described bag tegillum is coated on the micropore successively by mouse anti human albumin monoclonal antibody and bovine serum albumin(BSA) and forms.
2. people's denier albumin ELISA detection kit as claimed in claim 1, it is characterized in that: described solid phase carrier is 96 hole polystyrene agent plate.
3. people's denier albumin ELISA detection kit as claimed in claim 1, it is characterized in that: described solid phase carrier is 48 hole polystyrene agent plate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009201520440U CN201408194Y (en) | 2009-04-23 | 2009-04-23 | ELISA detection kit for trace albumin in human body |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009201520440U CN201408194Y (en) | 2009-04-23 | 2009-04-23 | ELISA detection kit for trace albumin in human body |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN201408194Y true CN201408194Y (en) | 2010-02-17 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009201520440U Expired - Fee Related CN201408194Y (en) | 2009-04-23 | 2009-04-23 | ELISA detection kit for trace albumin in human body |
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| Country | Link |
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| CN (1) | CN201408194Y (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102998464A (en) * | 2011-09-16 | 2013-03-27 | 武汉优尔生科技股份有限公司 | Dehydroepiandrosterone enzyme-linked immunosorbent assay kit development method |
| CN104407148A (en) * | 2014-09-04 | 2015-03-11 | 齐智 | Human albumin-supersensitive ELISA detection kit |
| CN108196043A (en) * | 2017-11-28 | 2018-06-22 | 泰州泽成生物技术有限公司 | Kit of microdose urine protein content and preparation method thereof in a kind of detection serum |
-
2009
- 2009-04-23 CN CN2009201520440U patent/CN201408194Y/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102998464A (en) * | 2011-09-16 | 2013-03-27 | 武汉优尔生科技股份有限公司 | Dehydroepiandrosterone enzyme-linked immunosorbent assay kit development method |
| CN104407148A (en) * | 2014-09-04 | 2015-03-11 | 齐智 | Human albumin-supersensitive ELISA detection kit |
| CN108196043A (en) * | 2017-11-28 | 2018-06-22 | 泰州泽成生物技术有限公司 | Kit of microdose urine protein content and preparation method thereof in a kind of detection serum |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20100217 Termination date: 20140423 |