CN2893705Y

CN2893705Y - Collaurum test paper for fast detecting shuangmeiwei residue (pesticide for treating downy mildew)

Info

Publication number: CN2893705Y
Application number: CN 200520142532
Authority: CN
Inventors: 万积成
Original assignee: Individual
Current assignee: Individual
Priority date: 2005-12-07
Filing date: 2005-12-07
Publication date: 2007-04-25
Anticipated expiration: 2015-12-07

Abstract

A colloidal gold test paper for fast detection of shuangmeiwei residue (pesticide for treating downy mildew), is composed of a base plate, a water absorption plate, a cellulose nitrate film, a monoclonal antibody gold marking pad, and a sample absorption layer. The cellulose nitrate film is placed in the middle of the base plate, and a shuangmeiwei synthetic immunogen test wire and a polyclonal antibody control wire are placed on the cellulose nitrate film. On one end of the base plate a water absorption plate is provided, and on the other end the sample absorption layer is positioned. The two ends of the cellulose nitrate film overlap and connect respectively with the water absorption plate and the monoclonal antibody gold marking pad. The sample absorption layer is pressed on the monoclonal antibody gold marking pad. The test paper is made using the colloidal gold immunity competition method which adopts the immunity colloidal gold method to detect whether the shuangmeiwei residue in fruits and vegetables exceeds specified standard. The method features in rapid, sensitive, simple and convenient operation which requires detection not necessarily by professional personnel, low cost which realizes the real unification of the complicated principle and simple operation, provides such virtues as easy storage and long validity terms.

Description

The colloid gold test paper that the fast detecting Propamocarb is residual

Technical field

The present invention relates to a kind of fruits and vegetables and raw grain based food Detecting Pesticide test paper and detection method thereof of being used for, belong to the colloid gold immune detection range.

Background technology

Because the vegetable growth phase is short and disease and pest is comparatively serious, thereby, in the growing vegetables process, usually need the continuous several times dispenser.Replace artificial weeding with the herbicide weeding in the presentization growing vegetables, raise labour efficiency.And many vegetables need repeatedly be gathered, gather and the time interval of dispenser short, cause when gathering the persticide residue in the vegetables often higher, be easy to surpass state specified standards.And persticide residue exceeds standard and can have a strong impact on level of human health in the vegetables, has become the food pollution source that can not be ignored.Government and each functional department extremely pay close attention to the residues of pesticides problem of food.But because the resistance to the action of a drug of the toxicity of agricultural chemicals and insect forms a vicious cycle, forbid thoroughly comprehensively that therefore the use of high highly toxic pesticide will have long process, even in using, low-toxin farm chemicals comprehensively, the pernicious poisoning of agricultural chemicals still takes place.Therefore new requirement and higher standard have all been proposed all many-sides such as the object of analyzing and testing, kind, quantity, scope, index.

Propamocarb (N-[3-(dimethylamino) propyl group] carbamic acid propyl ester hydrochloride), be a kind of new type bactericide of low toxicity, belong to carbamates.Its mechanism of action is to suppress the phosphatide of germ cell membrane component and the biosynthesizing of fatty acid, suppresses mycelial growth, sporangial formation and sprouting.The residual limit standard of regulation Propamocarb in the part agricultural product is in the residues of pesticides new standard of U.S.'s plan enforcement recently: leaf lettuce 65mg/kg, wheat wheat 0.05mg/kg, wheat straw 0.1mg/kg, wheat forage 0.3mg/kg, wheat hay 0.3mg/kg, cucurbit 1.5mg/kg, catsup 5mg/kg, fruit and vegetable 2mg/kg.Still do not have clear and definite standard and can follow though China limits the quantity of to Propamocarb in the crops residual, the U.S. is the main Agricultural Products Trade partner of China, and the plan of its residues of pesticides new standard is implemented the agricultural products in China outlet U.S. is produced certain influence.

At present, the analytical approach of detection residues of pesticides mainly is dependence, vapor-phase chromatography (GC) or gas chromatography and mass spectrum (MS) coupling method (GC/MS)." enzyme inhibition rate method+spectrophotometric method " has been listed in national proposed standard (GB/T 5009.199-2003).This method is quick, sensitive, easy and simple to handle and with low cost, can realize quick primary dcreening operation, but the restriction of examined scope and precision.And vapor-phase chromatography (GC) can realize the quantitative test of precision but speed is slower, and it is higher to detect cost.In a word, these method ubiquities the sample pretreatment process complexity, sensitivity be subjected to sample purification, step such as concentrate influence very big, consuming time, checkout equipment costliness, and require to have technical professional and long analytical cycle.

Therefore, utilize the competition law principle, on the basis of traditional immunodetection, introduced the collaurum Fast Detection Technique, develop Propamocarb collaurum competition law and detect test paper.This method has fast, sensitive, easy and simple to handle, cost is low, need not the professional detects, and really reaches the organic unity of complicated principle and ease of Use, have simultaneously preserve convenient, the characteristics that the term of validity is long.

Summary of the invention

The present invention is directed to some problems of above-mentioned existence, provide a kind of being applicable to detect the collaurum mensuration test paper whether Propamocarb exceeds standard.

For solving the problems of the technologies described above, the present invention is achieved by the following technical solutions:

At Propamocarb residue detection test paper base plate middle part is nitrocellulose filter, a test wire and a sheep anti mouse polyclonal antibody control line are arranged on the nitrocellulose filter, in base plate one end termination is water accepting layer, other end termination is the sample liquid-adsorption layer, the nitrocellulose filter two ends overlap mutually with Propamocarb monoclonal antibody gold mark pad with water accepting layer respectively and are connected (the overlapping coupling part is in 1-2 millimeter scope), are pressed with the sample liquid-adsorption layer on Propamocarb monoclonal antibody gold mark pad.Combined method about base plate, nitrocellulose filter, thieving paper, collaurum mark pad is carried out by patent CN 2741052Y.

The control line of Propamocarb residue detection test paper is formed by sheep anti mouse polyclonal antibody bag, and test wire is by Propamocarb synthetic immunogen bag quilt.The sample liquid-adsorption layer end that detects the residual colloid gold test paper of Propamocarb is put into Juice, because the capillarity sample will move along test strips water accepting layer end, when moving to Propamocarb monoclonal antibody gold mark pad, Propamocarb in the sample and Propamocarb monoclonal antibody gold mark probe generation specific bond, when moving to when being fixed with Propamocarb synthetic immunogen test wire, lose it and combine owing to Propamocarb in the antibody in the Propamocarb monoclonal antibody gold mark pad and the sample preferentially is combined into compound with the Propamocarb synthetic immunogen, therefore its collaurum can not be stranded on the test wire, the test wire place does not have red line to show, promptly has only a red control line positive; If do not have Propamocarb in the opposite sample, antibody in the Propamocarb monoclonal antibody gold mark pad moves on the test wire, the Propamocarb monoclonal antibody will with Propamocarb synthetic immunogen generation specific bond, collaurum is stranded on the test wire, promptly two red line are negative, immune competition law principle that Here it is.Propamocarb content just is inversely proportional in the shade of test wire and the sample.Carry out the transition to feminine gender from the positive, because the Propamocarb content difference of surveying, the color of test wire is also different, forms a red color gradient difference, detects the OD value with this principle, thereby infers the amount of actual Propamocarb that test wire reacts.

It detects more than the Propamocarb setting detected level to make test paper with competition law, and it is positive that a red line appears in the view window place; It is negative not decide the two red lines of the following appearance of detected level; Be that fuzzy hacures are boundary value at the test wire place when setting detected level, thereby draw judgement.

When moving to sheep anti mouse polyclonal antibody control line, no matter have or not Propamocarb in the sample, the gold mark probe of mark all can combine delay with the sheep anti mouse polyclonal antibody that has configured, and it is red that control line is shown.Therefore control line does not have colour band and produces that then the representative operation is wrong, during detection the sample liquid level surpass the MAX line or test paper expired.

Because adopt technique scheme, the residual colloid gold test paper of a kind of fast detecting Propamocarb provided by the present invention has such beneficial effect, promptly high specificity is highly sensitive, easily stores, need not the technical skill personnel operation, and readability as a result.

Description of drawings

Fig. 1 is the main TV structure figure of the residual colloid gold test paper of a kind of fast detecting Propamocarb of the present invention.

Fig. 2 is the side-looking structural drawing of invention figure one.

Fig. 3 shows positive findings figure for detecting.

Fig. 4 shows negative findings figure for detecting.

Among the figure 1, water sucting plate, 2, nitrocellulose filter, 3, sheep anti mouse polyclonal antibody control line, 4, test wire, 5, monoclonal antibody gold mark pad, 6, the sample liquid-adsorption layer, 7, base plate, 8, the MAX line.

Embodiment

1. according to the molecular structure of Propamocarb, infer the antigen determining area in its molecule, select for use the Propamocarb pesticide original medicine synthetic a kind of close with the Propamocarb molecular structure and have carboxyl or amino etc. and be easy to the be connected molecular structure (haptens) of group of protein macromolecule through chemical reaction.The haptens micromolecule is connected with protein macromolecule makes the Propamocarb immunizing antigen.

2. MONOCLONAL ANTIBODIES SPECIFIC FOR

(1) with synthetic immunogen immune BALB/c mouse.Set up the knurl strain with the SP2/0 Fusion of Cells and screen, get oncocyte and be injected in the BALB/c mouse abdominal cavity, make it produce ascites.

(2) extract the screening of mouse ascites purifying, the monoclonal antibody that acquisition can combine with the Propamocarb molecule is used for colloid gold label.

The preparation of collaurum and with the combining of Propamocarb monoclonal antibody

(1) get distilled water and add an amount of gold chloride magnetic agitation and be warmed to 85～95 ℃, add an amount of citrate three sodium and continue heated and stirred to seething with excitement 3～10 minutes, it is standby to keep in Dark Place after the cooling.

(2) the collaurum liquid of Propamocarb labeling of monoclonal antibody, on the adsorbing fiber material, dry back is standby.

4. film-making machine system film: utilize computer control transmission speed, guarantee that the antibody amount of bag quilt on the per unit film equates.

5. test strips combination: see Figure of description, method is with patent CN 2741052Y.

6. using method: the sample liquid-adsorption layer end of test paper is put into vegetables soak juice (liquid level must not surpass MAX line chart 8), take out test paper after 1 minute and keep flat, because kapillary and syphonic effect sample will move observations in the time of 5 minutes along test strips water accepting layer end.

7. the result judges: if the limit of identification of this test paper is set at 10mg/kg, then detect more than the Propamocarb concentration 10mg/kg, it is positive that a red line appears in the view window place; The two red lines of the following appearance of 10mg/kg are negative; Be that fuzzy hacures are boundary value at the test wire place during 10mg/g, this boundary value is the vegetable pesticide residue examination criteria 10mg/kg of country's promulgation.Thereby draw judgement.

Claims

1. the colloid gold test paper that the fast detecting Propamocarb is residual is characterized in that being made up of base plate (7), water sucting plate (1), nitrocellulose filter (2), Propamocarb monoclonal antibody gold mark pad (5), sample liquid-adsorption layer (6), MAX line (8); The base plate middle part is a nitrocellulose filter, a test wire (4) and a polyclonal antibody control line (3) are arranged on the nitrocellulose filter, base plate one end termination is a water sucting plate, other end termination is the sample liquid-adsorption layer, the nitrocellulose filter two ends overlap mutually with monoclonal antibody gold mark pad with water sucting plate respectively and are connected, and are pressed with the sample liquid-adsorption layer on monoclonal antibody gold mark pad.

2. the residual colloid gold test paper of a kind of fast detecting Propamocarb according to claim 1 is characterized in that control line is to be made into by sheep anti mouse polyclonal antibody bag.

3. the residual colloid gold test paper of a kind of fast detecting Propamocarb according to claim 1 is characterized in that test wire is to be made into by Propamocarb synthetic immunogen bag.

4. the residual colloid gold test paper of a kind of fast detecting Propamocarb according to claim 1 is characterized in that gold mark pad is as colloid gold label by the Propamocarb monoclonal antibody.

5. according to the residual colloid gold test paper of the described a kind of fast detecting Propamocarb of claim 1, it is Propamocarb 0.05～100mg/kg that the detected value that it is characterized in that test wire is set at the limit of identification scope.