CN209215397U - A kind of furazolidone fluorescence immunoassay immue quantitative detection reagent box - Google Patents

Abstract

The utility model discloses a kind of furazolidone fluorescence immunoassay immue quantitative detection reagent box, the kit includes the box cover (2) that box body (1) is flexibly connected therewith, is placed in box body (1) and examines 1 box of test strips, and 1 part of specification, 1, ID card, bar code 1 is opened；Agent plate (3) is additionally provided in box body (1) it is characterized in that, it is respectively arranged with the hole (4) for placing reagent bottle in agent plate (3) and places the groove (5) of reagent paper slip plastic clip shell, 4 holes (4) are divided into two rows of every row two and are distributed on agent plate (3), hole (4) is inner to placed 2 bottles of sample extracting solutions and 2 bottles of sample diluting liquids respectively, and hollow cylindrical drying device (6) are housed on hole (4).The kit is the kit of furazolidone content in the samples such as one kind at low cost, high sensitivity, high specificity, quick, accurate quantitative analysis detection animal muscle tissue, aquatic products and feed easy to operate.

Description

A kind of furazolidone fluorescence immunoassay immue quantitative detection reagent box

Technical field

The utility model belongs to detection kit field, and in particular to a kind of furazolidone fluorescence immunoassay quantitative detecting reagent Box.

Background technique

Furazolidone is itrofurans antimicrobial, is that the artificial synthesized wide spectrum begun to use in the later period forties in last century resists Bacterium drug can kill a variety of gram-positive bacterias and negative bacteria, be widely used in herding, aquaculture kind, but The residual of furazolidone and metabolin in animal derived food can pass to the mankind by food chain, and long-term intake can cause Various diseases have the side effects such as carcinogenic, teratogenicity to human body.The U.S., Japan and China have forbidden using in aquaculture The drug, but the drug is metabolized quickly in vivo, and since in November, 2005, China is repeatedly detected in the sea eel product of outlet Furazolidone metabolite product out, and people is caused to pay much attention to.

At present for furazolidone drug in animal derived food residue analysis method mainly have high performance liquid chromatography, LC-MS analysis method, enzyme-linked immunosorbent assay.High performance liquid chromatography, LC-MS analysis method need Want large-scale instrument and equipment, equipment price is expensive, and operating process is complicated, the time is longer, and can not at the scene with the small-sized reality in basis Test room use.Enzyme-linked immunosorbent assay is analyzed relative to instrument, easy to operate, at low cost, but accuracy is poor, is operated numerous It is trivial, it be easy to cause false positive or false negative etc..Therefore, it is quick to be badly in need of a kind of not only simplicity, but also the detection method of energy accurate quantitative analysis It detects the content of furazolidone in the samples such as animal muscle tissue, aquatic products and feed, meets scene i.e. quick and convenient and quasi- The requirement of true quantitative detection.

Summary of the invention

In order to overcome, existing method detection furazolidone is expensive, complicated for operation, detection time is long, can not on-site test Difficulty, solve the content of furazolidone in the samples such as real-time quick quantitative detection animal muscle tissue, aquatic products and feed.This Utility model provide it is a kind of quantify POCT detection kit using furazolidone time-resolved fluoroimmunoassay, kit use Reagent bottle mouth position install additional drying device, avoid detection environment in as humidity it is big caused by reagent in application and addition Be mixed into moisture and caused by accuracy reduce, especially extracting solution methylene chloride, since methylene chloride and water reaction can generate Poisonous gas causes adverse effect to human body, this kit can be effectively prevented this phenomenon, thus safer easy to be fast Speed, the content for detecting furazolidone drug in the samples such as animal muscle tissue, aquatic products and feed of accurate quantitative analysis.By immune The high degree of specificity of reaction and unique fluorescent microsphere are as antibody trace labelling object, and the time-resolved fluorescence with high sensitivity Immunochromatography technique and POCT theory combine, and it is wide, easy to operate to have invented a kind of high sensitivity, high specificity, the range of linearity Quick new detection kit meets the requirement of national basic veterinary drug residue detection.

To achieve the goals above, the utility model uses following technical scheme, and characterization step is as follows:

A kind of furazolidone fluorescence immunoassay immue quantitative detection reagent box, the kit include that box body (1) is flexibly connected therewith Box cover (2), interior be placed with of box body (1) examine 1 box of test strips, and 1 part of specification, 1, ID card, bar code 1 is opened；In box body (1) also Agent plate (3) is provided with it is characterized in that, having the hole (4) for placing reagent bottle in agent plate (3) and placing reagent paper slip The groove (5) that plastics get stuck, 4 holes (4) are divided into two rows of every row two and are distributed on agent plate (3), and hole (4) are inner to be placed respectively 2 bottles of sample extracting solutions and 2 bottles of sample diluting liquids are equipped with hollow cylindrical drying device (6) on hole (4).

It further illustrates, the drying device (6) is internal layer (S1) and two layers set of outer layer (S2) synthesis, one hollow circuit cylinder Shape structure is placed with desiccant in interlayer, and the internal layer of drying device (6) is provided with ventilation hole (7).

It further illustrates, is sealed above drying device (6) interlayer with sealant tape.

It further illustrates, drying device (6) outer layer is made of transparent material.

It further illustrates, the inspection test strips include: sample absorption pad, fluorescent microsphere conjugate pad, cellulose nitrate Plain reaction film, detection line, nature controlling line, water absorption pad and PVC plastic get stuck.

It further illustrates, plastics, which get stuck, is equipped with the well that sample absorption pad matches；Plastics, which get stuck, is additionally provided with nitric acid The fluorescence reading window that fibrin reaction film detection line and nitrocellulose reaction film nature controlling line match.

The beneficial effect comprise that:

(1) the utility model, which provides, a kind of quantifies POCT detection reagent using furazolidone time-resolved fluoroimmunoassay Box, overcome existing method detection furazolidone it is expensive, complicated for operation, detection time is long, can not on-site test difficulty, Solves the content of furazolidone in the samples such as real-time quick quantitative detection animal muscle tissue, aquatic products and feed.

(2) kit, which is used, installs drying device additional in reagent bottle mouth position, avoids in detection environment since humidity is big Caused by reagent be mixed into moisture in application and addition and caused by accuracy reduce.

(3) kit, which is used, installs drying device additional in reagent bottle mouth position, especially for extracting solution methylene chloride, due to Methylene chloride and water reaction can generate toxic gas, cause adverse effect to human body, this kit can be effectively prevented this Kind phenomenon.

In conclusion the kit easier quick, detection animal muscle tissue, aquatic products and feed of accurate quantitative analysis etc. The content of furazolidone drug in sample.By the high degree of specificity and unique fluorescent microsphere of immune response as antibody tracer Marker, and combined with highly sensitive time-resolved fluoroimmunoassay chromatographic technique and POCT theory, it has invented a kind of sensitive It spends that height, high specificity, the range of linearity be wide, simple and quick new detection kit, meets national basic veterinary drug residue detection and require.

Detailed description of the invention

Fig. 1 is kit structural map, including with lower component: 1, box body, 2, box cover (2), 3, agent plate, 4, hole, 5, groove, 6, drying device.

Fig. 2 is drying device sectional view, 7, venthole, S1 drying device outer layer, S2 drying device internal layer.

Fig. 3 fluorescence immunoassay quantitative test paper structural map, including with lower component: 1 ' sample pad, 2 ' bonding pads, 3 ' detection Line T, 4 ' nature controlling line C, 5 ' water absorption pads, 6 ' nitrocellulose filters, 7 ' .PVC plates, the furazolidone-embedded on 8 ' detection line T The monoclonal antibody of BSA antigen and the fluorescent microsphere reacted label, the goat-anti rabbit secondary antibody embedded on 9 ' nature controlling line C with And the fluorescent microsphere label rabbit igg antibody reacted on bonding pad.

Specific embodiment

The present invention will be illustrated by specific embodiment further progress below, such as in the whole text, specification and claim are worked as Mentioned in "comprising" or " comprising " be an open language, therefore should be construed to " including but not limited to ".Specification is subsequent Be described as implementing better embodiment of the invention, so it is described description be for the purpose of the rule of specification, not to It limits the scope of the invention.Protection scope of the present invention is as defined by the appended claims.

1. preparing furazolidone time-resolved fluoroimmunoassay quantifies POCT quick detection kit, kit includes following Content

Wherein test strip mainly got stuck by: PVC plastic, nitrocellulose filter, sample pad, bonding pad, T line, C line, Water absorption pad composition.

2. the assembling of furazolidone time-resolved fluoroimmunoassay quantitative testing test paper item: first using time resolution europium (Eu) fluorescence Microballoon marks furazolidone antibody, is then dried and is fixed on bonding pad, in combination with being also sprayed with time resolution Eu on pad The rabbit igg antibody of fluorescent microsphere label, then furazolidone-BSA antigen is fixed on nitrocellulose filter and is made into detection line T Line, while being coated with goat-anti rabbit secondary antibody on nitrocellulose filter and being made into Quality Control C line, 37 DEG C of drying are finally micro- by sample pad, fluorescence Ball conjugate pad, nitrocellulose filter and water absorption pad are successively overlapped in PVC board, then are cut into the test strips of 4mm wide.It is transferred to PVC Plastics get stuck, and plastics get stuck equipped with the well matched with the sample pad, and detect T line and Quality Control with the reaction film The fluorescence reading window that C line matches.

3. furazolidone time-resolved fluoroimmunoassay quantitative testing test paper test philosophy: adding when by sample to be tested dropwise addition When sample area, the furazolidone antibody knot merga pass capillary of fluorescent microsphere label in the furazolidone drug and bonding pad in sample Effect chromatographs forward, after reaching detection zone, fixed furazolidone antigen and remaining unbonded fluorescent microsphere on detection line T line Furazolidone antibody is marked to combine.In the furazolidone antibody concentration and sample of the fluorescent microsphere label combined on detection line T line The concentration of furazolidone is inversely proportional, and the fluorescent marker concentration that nature controlling line C line combines is unrelated with the concentration of furazolidone in sample. After chromatography, POCT analyzer is quantified using time-resolved fluoroimmunoassay and reads the fluorescence intensity of T line and the relative fluorescence of C line Intensity simultaneously calculates T/C value, by, with the ID card and bar code of standard curve and detection project information, being divided with the time in kit Distinguish that fluorescence immunoassay quantifies the concentration that the analysis software built in POCT analyzer calculates furazolidone in sample automatically.

4. it is dichloromethane that furazolidone time-resolved fluoroimmunoassay, which quantifies sample extracting solution in POCT quick detection kit, Alkane solution, sample diluting liquid are the 0.01mol/ containing 0.5% (m/v) bovine serum albumin(BSA), 2.5% (m/v) Tween20 The phosphate buffer solution of LpH7.0.Specification is the detailed description of testing principle, operating procedure, points for attention.ID card and bar shaped Code is with standard curve and detection project information, for the standard curve of automatic identification detection project and extraction this batch kit Information.

The innovation of the utility model is that building furazolidone time-resolved fluoroimmunoassay quantifies POCT test strip, and benefit Only divided with the unification platform time with time-resolved fluorescence immune quantitative POCT detection kit using the principle of competitive immunoreaction Distinguish fluorescence immunoassay quantify POCT analyzer, it can be achieved that convenient and efficient, inexpensive, highly sensitive quantitative detection animal muscle tissue, The content of furazolidone in the samples such as aquatic products and feed.Fluorescent marker is using lanthanide series Eu through special packet in test strips The fluorescent microsphere being process is wrapped up in, fluorescence intensity ratio general labeling object wants high 100 times or more, according to its luminous characteristics and Spectral Properties Property, transmitting fluorescence is measured with TIME RESOLVED TECHNIQUE, by time delay and wavelength resolution, effectively excludes the dry of non-specific fluorescence It disturbs, greatly improves sensitivity for analysis, keep the stability of the utility model kit test method and sensitivity better than existing There is detection method.With the 0.05 μ g/kg of sensitivity of the kit method detection furazolidone of the utility model, the range of linearity is 0.2-10 μ g/kg, accuracy TIANZHU XINGNAO Capsul are 80-125%, and 5 minutes accurate quantitative analysis of sample solution determine furazolidone medicine The content of object, testing result meet the U.S., in Chinese animal-derived food furazolidone residue limits.

Embodiment 1

The following are the detecting steps of the utility model kit detection furazolidone

1 experimental material and method

1.1 reagents and solution

The reagent solution of the utility model detection method is all contained in time-resolved fluoroimmunoassay and quantifies POCT detection reagent In box, it is not necessarily to other reagent preparation.

1.2 instrument and equipment

1.2.1 XT8201A type time-resolved fluoroimmunoassay quantifies POCT analyzer (the limited public affairs of Shanghai ambitious plan biotechnology Department provides)

1.2.2 XT8202A type constant-temperature incubation device

1.2.3 eddy blending machine

1.2.4 low speed centrifuge

1.2.5 electronic balance

1.2.6 homogenizer

1.3 test method

1.3.1 the pre-treatment of sample

1.3.1.1 the processing of animal muscle tissue, aquatic products and feed: by ground 40 mesh of feed, animal muscle group Knit and take edible meat about 200g with aquatic products, with homogenizer thoroughly it is homogeneous after, accurately weigh 2.0 ± 0.02g it is homogeneous after sample In 15ml centrifuge tube, sample extracting solution puts back to kit in the putting hole of drying device after opening lid, takes out 10ml Sample extracting solution is added in sample, with vortex oscillator mechanical shaking extraction 5min；After extraction, 4000rpm is centrifuged 2min, takes 100 μ l supernatants are added to 200 μ l sample diluting liquids, to sample detection after mixing.

1.3.2 detection process and result

1. constant-temperature incubation device is switched on, it is arranged 37 DEG C of temperature, rising to 37 DEG C of constant rears to temperature can be used；

2. being inserted into ID card corresponding to this batch products, and scan after fluorescence immunoassay quantifies POCT analyzer preheating 5min Bar code corresponding to this batch products；

3. taking out furazolidone time-resolved fluoroimmunoassay from aluminium foil bag quantifies POCT test strip, it is horizontally placed at perseverance On warm couveuse；

4. drawing sample to be tested of the 100 μ l Jing Guo pre-treatment with pipettor to be added in the well of test strips, incubation is covered Device lid starts timing and is incubated for 5min；

Test strips insertion time-resolved fluoroimmunoassay is quantified in POCT analyzer test strips slot after 5.5min, touch screen Point reading, reading value is the content of furazolidone in sample；

6. if had testing result exceeds the upper limit of the range of linearity by kit is put back to after sample diluting liquid opening lid In the putting hole of drying device, takes out sample diluting liquid and be added in sample, detected again after being diluted with sample diluting liquid, detection knot Fruit is multiplied by extension rate, the as ultimate density of sample.

Claims (5)

1. a kind of furazolidone fluorescence immunoassay immue quantitative detection reagent box, which includes the box that box body (1) is flexibly connected therewith It covers (2), is placed in box body (1) and examines 1 box of test strips, 1 part of specification, 1, ID card, bar code 1 is opened；It is also set in box body (1) It is equipped with agent plate (3), which is characterized in that have the hole (4) for placing reagent bottle in agent plate (3) and place reagent paper slip Groove (5), 4 holes (4) are divided into two rows, and every row two is distributed on agent plate (3), and hole (4) inner to be placed 2 bottles of samples respectively and mention Liquid and 2 bottles of sample diluting liquids are taken, hollow cylindrical drying device (6) are housed on hole (4), the drying device (6) is internal layer (S1) and two layers set of outer layer (S2) synthesis, one hollow columnar structures, be placed with desiccant in interlayer, drying device (6) it is interior Layer is provided with ventilation hole (7).

2. kit according to claim 1, which is characterized in that drying device (6) interlayer is sealed with sealant tape.

3. kit according to claim 1, which is characterized in that drying device (6) outer layer is made of transparent material.

4. kit described according to claim 1, which is characterized in that the inspection test strips include: sample absorption pad, glimmering Light microballoon conjugate pad, nitrocellulose reaction film, detection line, nature controlling line, water absorption pad and PVC plastic get stuck.

5. the kit according to claim 4, which is characterized in that plastics get stuck to be added equipped with what sample absorption pad matched Sample hole；Plastics, which get stuck, is additionally provided with nitrocellulose reaction film detection line and fluorescence that nitrocellulose reaction film nature controlling line matches Read window.