CN203519613U

CN203519613U - Test strip for semi-quantitative detection on 4-chlorophenoxyacetic acid

Info

Publication number: CN203519613U
Application number: CN201320384056.2U
Authority: CN
Inventors: 陈琰; 周俊; 冷毅斌
Original assignee: ELABSCIENCE BIOTECHNOLOGY Co Ltd
Current assignee: ELABSCIENCE BIOTECHNOLOGY Co Ltd
Priority date: 2013-06-28
Filing date: 2013-06-28
Publication date: 2014-04-02
Anticipated expiration: 2023-06-28

Abstract

The utility model discloses a test strip for semi-quantitative detection on a 4-chlorophenoxyacetic acid (4-D), which comprises a sample pad, a colloidal gold mat, a PVC (polyvinyl chloride) board, a nitrocellulose membrane and a water absorption pad, wherein the nitrocellulose membrane is adhered to the PVC board, the nitrocellulose membrane is provided with a detection area and a contrast area, the detection area is a detection line (T line), the contrast area is a quality control line (C line), the water absorption pad and the colloidal gold mat are adhered to the two end sides of the nitrocellulose membrane in a stacked mode, and the sample pad is adhered to the colloidal gold mat in a stacked mode. The test strip disclosed by the utility model has the beneficial effects that the test strip is simple in operation, quick in response, high in sensitivity, strong in specificity and suitable for field detection, and also has the advantages of short production cycle, high output, low cost and the like, therefore, the test strip has a good application prospect in the technical field of immune detection. The test strip is simple in operation, quick in response, high in sensitivity, strong in specificity and suitable for field detection, and also has the advantages of short production cycle, high output, low cost and the like.

Description

The test strips of half-quantitative detection 4-chlorophenoxyacetic acid

Technical field

The utility model belongs to immunochemistry detection technique, specifically, the test strips of a kind of half-quantitative detection 4-chlorophenoxyacetic acid (4-D), by the immunochromatography reaction of collaurum colour developing, to reach the object of the 4-chlorophenoxyacetic acid residues of pesticides in fast detecting agricultural product, water, soil.

Background technology

4-chlorophenoxyacetic acid is widely used herbicide and plant growth regulator on producing, it is at agricultural product, residual harm in water and soil can not be ignored, as it is at soil, residual exceeding standard in water, easily cause cotton, some vegetables, the mankind's health is caused to larger harm, it is reported, the carcinogenicity that 4-chlorophenoxyacetic acid tool is potential, mutagenicity, it is a kind of endocrine disruptor, to immune system and the harmful effect of fertility system tool, after human body take in to absorb to eyes, skin also has spread effect, repeatedly contact after to liver, heart has damage effect, can cause convulsions etc.Given this, some country and world organization have all made strict regulation to 4-chlorophenoxyacetic acid in environmental standard, the World Health Organization (WHO) promulgates < < drinking water quality guide > > in August, 2003, the maximum concentrations 30ug/l of 4-chlorophenoxyacetic acid in regulation potable water.The < < drinking water quality hygienic practice > > that China promulgates calendar year 2001 detects in project very much, by the restriction regulation of 4-chlorophenoxyacetic acid, be 0.03mg/l, to the conventional sense analysis of its detected level, can only be to utilize the instruments such as chromatography of gases (GC) and high performance liquid chromatography (HPLC) in laboratory operation, method is loaded down with trivial details, analysis speed is slow, cost is high, requires instrumentation degree high.Along with sample to be checked, particularly require field quick detection sample size and express delivery to increase, traditional residues of pesticides means are difficult to meet the need, therefore, and in the urgent need to Kaifeng and application high-level efficiency Analytical Techniques of Pesticide Residues.

By great mass of data, retrieve, notice: the preparation of 4-dichlorphenoxyacetic acid comlete antigen and antibody, and this artificial immunity antigen immune rabbit is prepared to polyclonal antibody, set up on this basis the technology of indirect ELISA detection 4-chlorophenoxyacetic acid residues of pesticides.This technology has larger innovation compared with GC, HPLC, make that detection time is shorter, cost is greatly lower, sensitivity improves, be conducive in food, soil, water conservancy environment, the fast detecting of 4-chlorophenoxyacetic acid residues of pesticides is also applied, but its method also exist operation also operate relatively loaded down with trivial details, need to repeatedly wash, need, by microplate reader reading etc., can not realize single stage method Fast Measurement truly.Also having biological heredity device is a kind ofly to take biologically active unit as sensitive element, in conjunction with chemistry, physical transformation element, instrument analyte to high selectivity, highly sensitive during pesticide residue analysis, this method can be brought into play great role in real-time test field, but because this method still needs certain instrument and equipment condition, is difficult for further reducing testing cost.

Colloidal gold immunochromatographimethod technology is grew up in the last few years a kind of simple, rapid free epidemiology detection method, tool is sensitive, simple, ripe, reliable without equipment and instrument, economic and practical, tool high degree of specificity and susceptibility, visual result, reagent and amount of samples seldom, do not need professional to operate soon, be easy to promote the use of, just can meet this demand.

Summary of the invention

The purpose of this utility model is to overcome deficiency of the prior art, the test strips of a kind of half-quantitative detection 4-chlorophenoxyacetic acid (4-D) is provided, change the loaded down with trivial details problem of 4-chlorophenoxyacetic acid residues of pesticides multi-step, realize cheap, step fast detecting 4-chlorophenoxyacetic acid residues of pesticides.

The utility model solves the problems of the technologies described above adopted technical scheme: the test strips of half-quantitative detection 4-chlorophenoxyacetic acid (4-D), it is characterized in that including sample pad, collaurum pad, PVC plate, nitrocellulose filter and adsorptive pads, wherein, on PVC plate, be pasted with nitrocellulose filter, on described nitrocellulose filter, there are detection zone and check plot, detection zone is detection line (T line), check plot is nature controlling line (C line), stacked adsorptive pads and the collaurum pad of being pasted with of both end sides of nitrocellulose filter, the stacked sample pad that is pasted with also on collaurum pad.

Press such scheme, described detection line (T line) 4 has four.

Press such scheme, described detection line (T line) is followed successively by T1 line, T2 line, T3 line, T4 line from left to right, in detection line (T line), 4-chlorobenzene oxy butyrate-BSA conjugate solution concentration increases progressively successively according to the order of T4 line, T3 line, T2 line, T1 line, and the total amount that is coated with variable concentrations 4-chlorobenzene oxy butyrate-BSA conjugate coating antigen just can with gold mark pad on golden labeling antibody neutralize completely, in order to detect 4-chlorophenoxyacetic acid (4-D) residues of pesticides of variable concentrations, check plot is coated with sheep anti-mouse igg polyclonal antibody.

It is haptens prepared by 4-chlorophenoxyacetic acid antibody that the utility model is selected 4-chlorobenzene oxy butyrate, and bovine serum albumin(BSA) (BSA) is carrier protein, then by the synthetic artificial antigen 4-chlorobenzene oxy butyrate-BSA of improvement EDC method.Take 4-chlorobenzene oxy butyrate 60mg, more than adding 0.05MPB(PH11), stirring is slowly dissolved it, regulate afterwards pH value to 7.2 left and right, add 60mg sulf-NHS, fully mix, add again 90mgEDC, stir evenly, standing a moment, and this reactant liquor is slowly added drop-wise in 1 hour and is prepared in advance in 60mg9mlBSA solution, and at 4 ℃, reaction stirred is spent the night, 0.15M NaCl dialysis three days, 5-6 left and right, ultraviolet spectrophotometer is surveyed the combination ratio of ultraviolet method measurement and calculation conjugate, obtain suitably in conjunction with than 4-chlorobenzene oxy butyrate-BSA conjugate, frozen at-20 ℃.

The beneficial effects of the utility model are: 4-chlorophenoxyacetic acid (4-D) detects reagent strip, simple to operate, reaction fast, high, the high specificity of susceptibility and applicable Site Detection, but also have with short production cycle, output is high, low cost and other advantages, in technical field of immunoassay, have a good application prospect.This test strips, simple to operate, reaction fast, high, the high specificity of susceptibility and applicable Site Detection, but also have with short production cycle, output is high, low cost and other advantages.

Accompanying drawing explanation

Fig. 1 structural drawing of the present utility model;

Fig. 2 vertical view of the present utility model;

Fig. 3 detects 4-chlorophenoxyacetic acid (4-chlorophenoxyacetic acid), and result is negative

Fig. 4 detects 4-chlorophenoxyacetic acid (4-chlorophenoxyacetic acid), and result is low content (20-100ng/ml)

Fig. 5 detects 4-chlorophenoxyacetic acid (4-chlorophenoxyacetic acid), and result is middle low content (100-1000ng/ml)

Fig. 6 detects 4-chlorophenoxyacetic acid (4-chlorophenoxyacetic acid), and result is middle content (1-10ug/ml)

Fig. 7 detects 4-chlorophenoxyacetic acid (4-chlorophenoxyacetic acid), and result is high-load (> 10ug/ml)

In figure, 1 is sample pad, and 2 is collaurum pad, and 3 is PVC plate, and 4 is p-wire (T line, totally 4), and 5 is nature controlling line (C line), and 6 is nitrocellulose filter, and 7 is adsorptive pads.

Embodiment

Below in conjunction with accompanying drawing and embodiment, the utility model is described in further detail.

Embodiment 1

As shown in Figure 1, the test strips of half-quantitative detection 4-chlorophenoxyacetic acid (4-D), it is characterized in that including sample pad 1, collaurum pad 2, PVC plate 3, nitrocellulose filter 6 and adsorptive pads 7, wherein, on PVC plate 3, be pasted with nitrocellulose filter 6, on described nitrocellulose filter, have detection zone and check plot, there are four detection lines (T line) 4 detection zone, and check plot is nature controlling line (C line) 5, stacked adsorptive pads 7 and the collaurum pad 2 of being pasted with of both end sides of nitrocellulose filter 6, the stacked sample pad 1 that is pasted with also on collaurum pad 2.

Described detection line (T line) is followed successively by T1 line, T2 line, T3 line, T4 line from left to right, in detection line (T line), 4-chlorobenzene oxy butyrate-BSA conjugate solution concentration increases progressively successively according to the order of T4 line, T3 line, T2 line, T1 line, and be coated with variable concentrations 4-chlorobenzene oxy butyrate-BSA conjugate coating antigen, on T line detection line the total amount of coating antigen just can with gold mark pad on golden labeling antibody neutralize completely, in order to detect 4-chlorophenoxyacetic acid (4-D) residues of pesticides of variable concentrations, check plot is coated with sheep anti-mouse igg polyclonal antibody.

According to competition Immunosuppression chromatographic theory, the coated variable concentrations 4-chlorobenzene oxy butyrate-BSA conjugate in detection zone on nitrocellulose filter 6, check plot is coated with sheep anti mouse polyclonal antibody.While not containing 4-chlorophenoxyacetic acid in sample, the amount of the anti-4-chlorophenoxyacetic acid antibody of collaurum on glass fibre element film just can make 4 detection lines colour developings on nitrocellulose filter, and nature controlling line also develops the color; While having tested antigenic substance (4-chlorobenzene oxy butyrate) in determinand, these residues of pesticides will form Ag-Ab-Au compound with the anti-4-chlorobenzene oxy butyrate antibody of mark collaurum, and the medicine antigen that in this compound, the avtive spot of anti-4-chlorobenzene oxy butyrate antibody occupies the left drug because of by sample solution on cannot the detection line on detection zone is combined.

During working sample, test strips absorption of sample pad is inserted in testing sample, stop and take out and keep flat, observations after 10 minutes afterwards for 10 seconds.Result determination methods is by the band quantity on demonstration and immunochromatography film, judges the content that detects 4-chlorophenoxyacetic acid in sample.So when in sample, 4-chlorophenoxyacetic acid is low content (20-100ng/ml),

detection line

1,2,3 and nature controlling line colour developing, detection line 4 does not develop the color; When in sample, 4-chlorophenoxyacetic acid is middle low content (100-1000ng/ml),

detection line

1,2 and nature controlling line colour developing, the 3rd, 4 detection lines do not develop the color; When in sample, 4-chlorophenoxyacetic acid is middle content (1-10ug/ml), detection line 1 and nature controlling line colour developing,

detection line

2,3,4 does not develop the color; When in sample, 4-chlorophenoxyacetic acid is high-load (> 10ug/ml),

detection line

1,2,3,4 does not all develop the color, and a nature controlling line colour developing, is judged to be feminine gender.This detection completed in 10 minutes, simple to operate, reaction fast, high, the high specificity of susceptibility and applicable Site Detection, but also have with short production cycle, output is high, low cost and other advantages.In technical field of immunoassay, have a good application prospect.

In a word, the foregoing is only preferred embodiment of the present utility model, all equalizations of doing according to the utility model claim change and modify, and all should belong to the covering scope of the utility model patent.

Claims

1. the test strips of half-quantitative detection 4-chlorophenoxyacetic acid, it is characterized in that including sample pad (1), collaurum pad (2), PVC plate (3), nitrocellulose filter (6) and adsorptive pads (7), wherein, on PVC plate (3), be pasted with nitrocellulose filter (6), on described nitrocellulose filter, there are detection zone and check plot, detection zone is detection line (4), check plot is nature controlling line (5), stacked adsorptive pads (7) and the collaurum pad (2) of being pasted with of both end sides of nitrocellulose filter (6), the upper also stacked sample pad (1) that is pasted with of collaurum pad (2).

2. by the test strips of half-quantitative detection 4-chlorophenoxyacetic acid claimed in claim 1, it is characterized in that described detection line (4) has four.