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Immunological detection method and kit for organophosphorus and carbamate pesticides

A technology of carbamates and organic phosphorus, applied in the direction of measuring devices, scientific instruments, instruments, etc., to achieve the effects of low cost, wide detection range, and cost-saving detection

Active Publication Date: 2019-08-30
赵芳
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, both the ELISA substance and the immunochromatographic test paper method can only detect one or two (five or six) pesticide residues at a time, which cannot meet the actual needs of hundreds of pesticides currently in use.

Method used

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  • Immunological detection method and kit for organophosphorus and carbamate pesticides
  • Immunological detection method and kit for organophosphorus and carbamate pesticides
  • Immunological detection method and kit for organophosphorus and carbamate pesticides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Processing method of samples to be tested

[0020] 1. Meat: 1) Take 0.5g of meat in a 10ml beaker, add 5ml of acetone to soak for 5min, shake once every 1min, then add 0.2g of calcium carbonate; 2) Take 0.5ml of the above liquid in a 5ml beaker, Blow dry acetone; add 0.3ml 0.1M Tris-HCl (pH8.0) buffer solution to dissolve; 3) add 0.1ml 0.3% calcium hypochlorite, let stand for 10min; 4) add 0.3ml 10% sodium nitrite, shake well , for future inspection.

[0021] 2. Milk: 1) Raw milk and pasteurized milk can be tested directly without any treatment. 2) Fermented milk: adjust the pH to pH8.0 with twice the mass of 0.1MTris-HCl (pH8.0) for inspection. 3) Milk powder: Take 1g of milk powder, add 9ml of deionized water, dissolve, and prepare for inspection.

[0022] 3. Honey: dilute 10 times with 0.1M Tris-HCl (pH8.0) for inspection.

[0023] 4. Urine: Dilute 2 times with 0.1M Tris-HCl (pH8.0) for inspection.

[0024] 5. Fruits and vegetables: 1) Take 1g vegetabl...

Embodiment 2

[0027] Example 2 The immunochromatographic method for detecting DTNB (substance D)

[0028] 1. Preparation of gold-labeled anti-5,5-dinitro-bis(2-nitrobenzoic acid) (DTNB) monoclonal antibody: take 10ml of gold nanoparticles with a particle size of 40nm, add 10μg anti-DTNB monoclonal antibody; room temperature React for 15 minutes; add BSA to a final concentration of 1% (W / V), and continue to react at room temperature for 15 minutes; centrifuge, and resuspend the particles with 1ml of reconstitution solution (10mM Tris-HCl (pH8.0), 0.5%BSA), and set aside.

[0029] 2. Preparation of chromatographic membrane: Spray DTNB-carrier protein conjugates with a concentration of 2 mg / ml on the Sartorius CN95 nitrocellulose membrane in an amount of 0.8 μl / cm (DTNB detection area, which can be DTNB-cow Serum albumin, DTNB-ovalbumin or covalent conjugates of DTNB and other protein molecules), 0.05mg / ml goat anti-mouse IgG (quality control area), dry at 37°C for 2h, and set aside.

[0030]...

Embodiment 3

[0051] Example 3 The immunochromatographic method for detecting TNB (substance C)

[0052] 1. Preparation of gold-labeled anti-5-mercapto-2-nitrobenzoic acid (TNB): take 10ml of gold nanoparticles with a particle size of 40nm, add 13μg of anti-TNB monoclonal antibody; react at room temperature for 15min; add BSA to a final concentration of 1% (W / V), continue to react at room temperature for 15 minutes; centrifuge, resuspend the particles with 1ml reconstitution solution (10mM Tris-HCl (pH8.0), 0.5%BSA), and set aside.

[0053] 2. Preparation of chromatographic membrane: Spray TNB-carrier protein conjugates with a concentration of 2mg / ml on the Sartorius CN95 nitrocellulose membrane sequentially according to the amount of 1.0μl / cm (TNB detection area, which can be TNB-bovine Serum albumin, TNB-ovalbumin or covalent conjugates of TNB and other protein molecules), 0.03mg / ml goat anti-mouse IgG (quality control area), dry at 37°C for 2h, and set aside.

[0054] 3. Preparation of ...

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Abstract

The invention discloses an immunological detection method for organic phosphorus and carbamates pesticides. An immunological method is adopted for detecting the change condition of cholinesterase substrate so as to judge if a sample contains the organic phosphorus and carbamates pesticides. The invention also discloses a kit for detecting the organic phosphorus and carbamates pesticides. According to the invention, the residual condition of the organic phosphorus and carbamates pesticides in the detected sample can be judged in the manner of indirectly detecting the activity of cholinesterase. The method has the advantages of wide detection scope, high sensitivity, being simple and feasible and low cost.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to an immunological detection method and kit for organophosphorus and carbamate pesticides. Background technique [0002] Organophosphorus and carbamate pesticides have been widely used in my country's agricultural production practice due to their broad insecticidal spectrum and high efficacy. Due to its large usage and wide range of applications, it has received widespread attention in terms of food safety and environmental protection. In particular, some highly toxic and highly toxic pesticides, such as methamidophos, methyl parathion, parathion and other pesticides, have been banned from being used on vegetables and fruits due to their acute toxicity and residual toxicity. However, due to the lack of scientific knowledge of pesticides used by farmers, the problem of indiscriminate use and abuse of highly toxic and highly toxic pesticides often occurs. Even for those pesticid...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/573G01N33/543
Inventor 赵芳史跃光其他发明人请求不公开姓名
Owner 赵芳
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