CN201984069U

CN201984069U - Melamine luminous ELIsA (enzyme linked immunosorbent assay) kit

Info

Publication number: CN201984069U
Application number: CN2011200737541U
Authority: CN
Inventors: 张宏斌
Original assignee: GUANGZHOU JIHENG MEDICAL TECHNOLOGY Co Ltd
Current assignee: GUANGZHOU JIHENG MEDICAL TECHNOLOGY Co Ltd
Priority date: 2011-03-18
Filing date: 2011-03-18
Publication date: 2011-09-21
Anticipated expiration: 2021-03-18

Abstract

A melamine luminous ELIsA kit aims to provide a kit with the advantages of low use cost, convenience for operation, high sensitivity and wide application range, and belongs to the technical field of food chemistry detection. The utility model adopts the technical keys that 14 reagent bottles, an enzyme-linked plate, a group of reagent bottle racks and an enzyme-linked plate lower groove are arranged in the kit; the reagent bottles are placed on the reagent bottle rack; the enzyme-linked plate is placed in the enzyme-linked plate lower groove; and a cover plate film is covered on the enzyme-linked plate. The kit can be used for detecting the melamine content in food.

Description

The luminous enzyme-linked immuno assay kit of a kind of melamine

Technical field

The utility model relates to a kind of kit, particularly relates to the luminous enzyme-linked immuno assay kit of a kind of melamine.

Background technology

Melamine relates to harmful small organic molecule matter of food security, after entering human body, substitution reaction (hydrolysis) can take place, generate cyanuric acid, cyanuric acid and melamine form big reticulate texture, cause calculosis, but some manufacturers add melamine in violation of rules and regulations for protein content index in the raising milk powder, as the Sanlu milk powder case.

At present melamine detection being relied on high performance liquid chromatography and liquid chromatography-mass spectrography and minority need be from the detection platform of external import, the pre-treatment process complexity of these detection methods, detection time is long, the testing cost height, instrument and equipment is more expensive and lack defective such as independent intellectual property right and seriously restricted China to the harmful micromolecular effective detection of food security, has endangered broad masses of the people's the healthy and correlation detection technology and the development of industry.

Therefore, utilize advanced technology platform, set up detectable and method with independent intellectual property right, solve the harmful micromolecule of the food security that has serious harm at present detect difficult and multiple detectable and method to rely on external import problem extremely urgent.

The utility model content

At above-mentioned deficiency, the utility model provides that a kind of use cost is low, easy to operate, the luminous enzyme-linked immuno assay kit of content of melamine in the fast detecting food.

The technical solution of the utility model is such: the luminous enzyme-linked immuno assay kit of a kind of melamine, comprise box body, be provided with 14 reagent bottles, 1 elisa plate, cover plate film, a group reagent bottle stand and an elisa plate low groove in the box body, reagent bottle is placed on the reagent bottle stand, elisa plate is placed in the elisa plate low groove, is stamped the cover plate film on the elisa plate.

The luminous enzyme-linked immuno assay kit of above-mentioned a kind of melamine, wherein, described elisa plate low groove is arranged on the left end or the right-hand member of reagent bottle stand.

The luminous enzyme-linked immuno assay kit of above-mentioned a kind of melamine, wherein, described elisa plate is dismountable 96 hole elisa plates, bag is made solid phase antigen by the melamine with bovine serum albumin(BSA) lotus root connection on the micropore of elisa plate.

Further, the luminous enzyme-linked immuno assay kit of above-mentioned a kind of melamine, wherein, described 14 reagent bottles are respectively: two bottledly have a sample standard product reagent bottle, and six bottledly have a standard solution reagent bottle, one bottledly has a sample diluting liquid reagent bottle, one bottle is detected the solution A reagent bottle, a bottled detection solution B reagent bottle, and one bottledly has a washing stoste reagent bottle, the one bottled reagent bottle that substrate solution is arranged, one bottledly has a stop buffer reagent bottle.

Further, the luminous enzyme-linked immuno assay kit of above-mentioned a kind of melamine, wherein, described 14 reagent bottles are respectively: two bottledly have sample standard product reagent bottle, a 1ml; Six bottledly have standard solution reagent bottle, a 3ml; One bottledly has sample diluting liquid reagent bottle, a 20ml; One bottle is detected solution A reagent bottle, 1ml; The reagent bottle of one bottled detection solution B, 1ml; One bottledly has washing stoste reagent bottle, a 30ml; One bottledly has substrate solution reagent bottle, a 10ml; One bottledly has stop buffer reagent bottle, a 10ml.

The luminous enzyme-linked immuno assay kit of above-mentioned a kind of melamine, wherein, described box body is the hard box body.

Compared with prior art, the utlity model has advantages such as use cost is low, easy to operate, highly sensitive, applied range.

Description of drawings

Be further described below in conjunction with the drawings and specific embodiments kit of the present utility model, but do not constitute any restriction of the present utility model.

Fig. 1 is the utility model kit synoptic diagram;

Fig. 2 is that the utility model is opened synoptic diagram;

Fig. 3 is a reagent rack synoptic diagram of the present utility model;

Fig. 3 is the synoptic diagram of elisa plate of the present utility model.

Wherein: box body 1, reagent bottle stand 2, sample standard product reagent bottle 3, standard solution reagent bottle 4, sample diluting liquid reagent bottle 5 detects solution A reagent bottle 6, detects solution B reagent bottle 7, washing stoste reagent bottle 8, substrate solution reagent bottle 9, stop buffer reagent bottle 10, elisa plate 11, elisa plate low groove 12, cover plate film 13.

Embodiment

Embodiment 1

As shown in Figures 1 to 3, the luminous enzyme-linked immuno assay kit of a kind of melamine of the utility model, comprise box body 1, be provided with reagent bottle stand 2 in the box body 1, being placed with 2 on the reagent bottle stand 2 bottledly has a sample standard product reagent bottle 3, volume is 1ml, 6 bottledly have a standard solution reagent bottle 4, and volume is 3ml, and 1 bottledly has a sample diluting liquid reagent bottle 5, volume is 20ml, 1 bottle is detected solution A reagent bottle 6, and volume is 1ml, 1 bottled detection solution B reagent bottle 7, volume is 1ml, 1 bottledly has a washing stoste reagent bottle 8, and volume is 30ml, and 1 bottledly has a substrate solution reagent bottle 9, volume is 10ml, 1 bottledly has a stop buffer reagent bottle 10, and volume is 10ml, and the left end of reagent bottle stand is provided with the elisa plate low groove 12 of placing elisa plate 11, wherein, elisa plate is 96 holes, and bag is made solid phase antigen by the melamine with bovine serum albumin(BSA) lotus root connection on the micropore of elisa plate, elisa plate low groove top is provided with a cover plate film 13, is not subjected to the influence of introduced contaminants to guarantee elisa plate.

Experimental principle: this kit is used content of melamine in the luminous enzyme-labeled immunity assay sample.With the melamine bag of bovine serum albumin(BSA) (BSA) lotus root connection by microwell plate, make solid phase antigen, add melamine sample to be checked successively in the micropore of bag quilt, the anticalin molecular criteria product that can combine, the antiphagin antibody of HRP mark with the melamine specificity, through thorough washing back developing the color with the substrate luminol.Luminol generates the excited state intermediate under the catalysis of peroxidase, luminous when it gets back to ground state, the melamine in light intensity and the sample is negative correlation, and it detects wavelength is 425nm, and the luminescence immunoassay instrument detects, calculation sample concentration.

Kit is formed and the reagent preparation:

1. 1: one of elisa plate (96 holes, detachable).

2. standard items (dried frozen aquatic products): 2 bottles, every bottle is faced with preceding and is diluted to 1ml with sample diluting liquid, fully dissolving, its concentration is 1mg/L, does serial doubling dilution again, and dilution is 1000 μ g/L respectively, 500 μ g/L, 250 μ g/L, 125 μ g/L, 62.5 μ g/L, 31.2 μ g/L, 15.6 μ g/L, 0 μ g/L, stoste is directly as normal concentration 1000 μ g/L, and sample diluting liquid 3 is directly as normal concentration 0 μ g/L.

3. sample diluting liquid: 1 bottle, the 20ml/ bottle.

4. detection solution A: 1 bottle, 100 μ l/ bottles.Face with preceding and dilute by 1: 1000 to detect dilution, basis is calculated the required total amount configuration of each experiment well in advance before the dilution.

5. detection solution B: 1 bottle, 100 μ l/ bottles.Face with preceding and dilute by 1: 1000 to detect dilution, basis is calculated the required total amount configuration of each experiment well in advance before the dilution.

6. detection dilution: 2 bottles, the 10ml/ bottle.

7. washing stoste: 1 bottle, the 30ml/ bottle.Use 25 times of distilled water dilutings during use.

8. substrate solution: 1 bottle, the 10ml/ bottle.

9. stop buffer: 1 bottle, the 10ml/ bottle.

Wherein:

Described sample diluting liquid comprises phosphate buffer;

Described detection solution A is anticalin;

Described detection solution B is the antiphagin antibody of horseradish peroxidase HRP mark;

Described washing stoste comprises phosphate buffer;

Described substrate solution is a luminol solution;

Described stop buffer is a sulfuric acid solution;

Described elisa plate is 96 hole elisa plates.

The collection of sample and preservation:

Gather food samples, fully dissolve with sample diluting liquid in the drying tube, centrifugal 10 minutes of room temperature 1000g gets supernatant and can detect, or sample-20 a ℃ preservation is detected later on, but should avoid multigelation.

Operation steps:

All ingredients before use balance to room temperature.

1. application of sample: establish blank well, gauge orifice, testing sample hole respectively.Except that blank well, surplus hole adds standard solution or testing sample 100ul respectively.

2. need not wash, every hole adds detects solution A working fluid 100ul, 37 ℃, light shaking 60 minutes.

3. the washing working fluid is washed plate 3 times, and each every hole of 350ul/ dries.

4. every hole adds detects solution B working fluid 100ul, and 37 ℃, 60 minutes, wash plate 5 times, dry.

5. every in regular turn hole adds substrate solution 90ul, and 37 ℃ of lucifuges developed the color 10 minutes.

6. every in regular turn hole adds stop bath 50ul, cessation reaction.The luminescence immunoassay instrument is in each hole luminous intensity of 425nm wavelength measurement.

Specificity: this kit can detect each somatotype of melamine simultaneously, and with other albumen no cross reactions.

Calculate: the concentration with reference material is horizontal ordinate (logarithmic coordinate), and light intensity is ordinate (a common coordinate), draws typical curve on semilogarithmic paper, and light intensity value is per sample found corresponding concentration by typical curve, multiply by extension rate again; Or the linear regression equation that calculates typical curve with the concentration and the light intensity value of reference material, the light intensity value substitution equation with sample calculates sample concentration, multiply by extension rate again, is the actual concentrations of sample.

Sensing range: 15 μ g/L～1000 μ g/L

The utility model is not limited to above embodiment, so long as the scheme of mentioning in this specification and claims book all can be implemented.

Claims

1. luminous enzyme-linked immuno assay kit of melamine, comprise box body (1), it is characterized in that, be provided with 14 reagent bottles, an elisa plate (11), a group reagent bottle stand (2) and an elisa plate low groove (12) in the box body (1), reagent bottle is placed on the reagent bottle stand (2), elisa plate (11) is placed in the elisa plate low groove (12), is stamped cover plate film (13) on the elisa plate (11).

2. the luminous enzyme-linked immuno assay kit of a kind of melamine according to claim 1 is characterized in that described elisa plate low groove (12) is arranged on the left end or the right-hand member of reagent bottle stand.

3. the luminous enzyme-linked immuno assay kit of a kind of melamine according to claim 1, it is characterized in that, described elisa plate is dismountable 96 hole elisa plates, and bag is made solid phase antigen by the melamine with bovine serum albumin(BSA) lotus root connection on the micropore of elisa plate (11).

4. the luminous enzyme-linked immuno assay kit of a kind of melamine according to claim 1, it is characterized in that, described 14 reagent bottles are respectively: two bottledly have a sample standard product reagent bottle (3), six bottledly have a standard solution reagent bottle (4), one bottledly has a sample diluting liquid reagent bottle (5), one bottle is detected solution A reagent bottle (6), one bottled detection solution B reagent bottle (7), one bottled have the washing stoste reagent bottle (8), one bottledly has a substrate solution reagent bottle (9), and one bottledly has a stop buffer reagent bottle (10).

5. the luminous enzyme-linked immuno assay kit of a kind of melamine according to claim 4, it is characterized in that, described 14 reagent bottles are respectively: two bottledly have a sample standard product reagent bottle (3), 1ml, six bottledly have standard solution reagent bottle (4), a 3ml, one bottledly has a sample diluting liquid reagent bottle (5), 20ml, one bottle is detected solution A reagent bottle (6), 1ml, one bottled detection solution B reagent bottle (7), 1ml, one bottledly has washing stoste reagent bottle, a 30ml, one bottledly has a substrate solution reagent bottle, 10ml, one bottledly has stop buffer reagent bottle, a 10ml.

6. the luminous enzyme-linked immuno assay kit of a kind of melamine according to claim 1 is characterized in that described box body (1) is the hard box body.