Immune colloidal gold test strip for detecting organophosphorus pesticide and method for making same

A technology of organophosphorus pesticides and colloidal gold, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the requirements that cannot be used to detect organophosphorus pesticide residues in vegetables and fruits, cannot guarantee food quality and safety, and cannot meet timely and rapid detection, etc. problems, to achieve the effect of reliable detection results, easy promotion and application, and image judgment

Inactive Publication Date: 2007-09-26
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] At present, the main methods for the detection of pesticide residues in my country still rely on liquid chromatography (HPLC), gas chromatography (GC), liquid chromatography-tandem mass spectrometry (HPLC/MS/MS), gas chromatography-mass spectrometry (GC/MS). ) and other large-scale modern analytical instruments. Although these instruments have high detection sensitivity, they are expensive and require highly trained professional technicians to operate and use. At the same time, it takes several hours or even overnight from sample processing to final results. The operation process during this period requires several steps to more than ten steps, which cannot meet the timely and rapid detection of large-scale samples or temporary samples technically and economically, so that people's requirements for food quality and safety in daily life cannot be guaranteed. Therefore, research on vegetable And the rapid detection technology of pesticide residues in food has become a concern of the competent departments at all levels and the general residents. Therefore, sensitive, rapid and accurate detection equipment and methods for organophosphorus should be developed as soon as possible for the detection of vegetables, fruits, tissues and other samples. Organophosphorus residues are of great signific

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  • Immune colloidal gold test strip for detecting organophosphorus pesticide and method for making same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1: (detect the immune colloidal gold test strip of organophosphorus pesticide)

[0025] An immune colloidal gold test strip for detecting organophosphorus pesticides, the test strip includes a PVC backing 1, a self-adhesive adhesive layer 2, a polyester fiber sample pad 3, and is coated with an anti-organophosphorus pesticide-specific monoclonal antibody- -Colloidal gold marker polyester fiber binding pad 4, nitrocellulose membrane 5, detection zone coated with organophosphorus pesticide-BSA conjugate 7, control zone coated with goat or rabbit anti-mouse IgG and filter paper Water-absorbing pad 10; wherein, the detection band 7 is arranged on the left side of the middle end of the nitrocellulose membrane 5, and the contrast band 8 is arranged on the right side of the middle end of the nitrocellulose membrane 5, with an interval of 8-10 mm between the two. A limit line 11 is set on the top surface of the sample pad 3 corresponding to the left end of the bond p...

Embodiment 2

[0026] Embodiment 2: (detect the preparation method 1 of organophosphorus pesticide immune colloidal gold test strip)

[0027] Prepare as follows:

[0028] 1) Preparation of organophosphorus pesticide-BSA carrier protein conjugates: mix organophosphorus pesticides and BSA bovine serum albumin conjugates at a ratio of 1:5 (mol / mol), so that organophosphorus pesticides and BSA form stable particles , forming an organophosphorus pesticide-BSA conjugate through purification;

[0029] 2) Preparation of specific monoclonal antibodies against organophosphorus pesticides: immunize BALB / c mice with 50ug-100ug / carrier protein coupled with organophosphate three times, each time at an interval of 15-30 days; after the third booster immunization On the 3rd day, the eyes of the immunized mice were bled, and the neck was pulled to death, soaked in 75% alcohol for 5-10 minutes, the spleen was aseptically removed, crushed, filtered through a 100-mesh nylon mesh, and centrifuged at 1000r / min f...

Embodiment 3

[0035] Embodiment 3: (detect the preparation method 2 of organophosphorus pesticide immune colloidal gold test strip)

[0036] In this example, the organophosphorus pesticide and the BSA carrier protein were mixed at a ratio of 1:25 (mol / mol) to form an organophosphorus pesticide-BSA conjugate; the hybridoma cells were cultured in vitro to collect the culture supernatant Anti-organophosphorus pesticide-specific monoclonal antibody was prepared in liquid way; mouse antiserum was extracted to immunize rabbits, and rabbit anti-mouse IgG was obtained after purification; chloroauric acid was reduced to 30nm colloidal gold particles and mixed with 1:0.010 (ml / mg), the organophosphorus monoclonal antibody was added to the colloidal gold sol at pH 8.5-9.5 to form a stable colloidal gold complex, and the rest of the preparation methods were the same as in Example 2.

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Abstract

This invention discloses one immune glue gold test bar and process method for organophosphorus in agriculture test technique field, wherein, the test bar comprises PVC underlay, adhesive agent layer, sample pad, combination pad, aqua fortis fiber film, weight band, test band, compare band, overlap band, water absorptive pad, limit line and arrow line. The process method comprises the following steps: processing the medicine BSA load protein coupler; processing the medicine special signal clone antigen; processing sheep, rabbit or mouse antigen IgG; processing glue gold; processing single clone antigen gold label.

Description

technical field [0001] The invention relates to the technical field of pesticide detection, in particular to a test strip for detecting organophosphorus pesticide residues using antibody immune colloidal gold technology and a preparation method thereof. Background technique [0002] At present, the main methods for the detection of pesticide residues in my country still rely on liquid chromatography (HPLC), gas chromatography (GC), liquid chromatography-tandem mass spectrometry (HPLC / MS / MS), gas chromatography-mass spectrometry (GC / MS). ) and other large-scale modern analytical instruments. Although these instruments have high detection sensitivity, they are expensive and require highly trained professional technicians to operate and use. At the same time, it takes several hours or even overnight from sample processing to final results. The operation process during this period requires several steps to more than ten steps, which cannot meet the timely an...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/558G01N33/544
Inventor 朱亚红
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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