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Detection preparation for detecting atrazine, kit and detection method thereof

A technology of atrazine and a kit, which is applied in the field of detection preparations for detection of atrazine, can solve problems such as influence, and achieve the effects of wide linear range, low detection limit and high sensitivity

Pending Publication Date: 2021-12-21
INST OF ENVIRONMENTAL MEDICINE & OCCUPATIONAL MEDICINE ACAD OF MILITARY MEDICINE ACAD OF MILITARY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the traditional colloidal gold immunochromatographic analysis methods using naked eyes are mostly semi-quantitative detection, and different operators and interference factors are likely to have a greater impact on the test results

Method used

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  • Detection preparation for detecting atrazine, kit and detection method thereof
  • Detection preparation for detecting atrazine, kit and detection method thereof
  • Detection preparation for detecting atrazine, kit and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Step 1 Preparation of colloidal gold nanoparticles

[0069] Soak the stirring paddle and glassware in the acid tank overnight and clean; assemble the stirring paddle to the stirrer, turn on the electric heating mantle to preheat for 5 minutes; after the water in the flask boils, add 1mL of 1% sodium citrate, and turn on the Mix it evenly; adjust the rotation speed to 500rpm, and quickly add 1mL of 1% chloroauric acid. After the solution turns wine red, remove the heat source and continue to stir; stop stirring when the temperature drops to about 50°C, take the colloidal gold and use a volumetric flask to set the volume to 100mL, and measure the ultraviolet absorption peak.

[0070] Step 2 Preparation and optimization of atrazine monoclonal antibody-colloidal gold label

[0071] Take 1 mL of colloidal gold with OD520=1, add 20 μL of 0.1M sodium carbonate to adjust the pH to about 8.5, add 7 μL of 1 mg / mL atrazine, mix for 30 minutes, add 120 μL of 10% BSA, block for 15 ...

Embodiment 2

[0073] The preparation of embodiment 2 test strips

[0074] Preparation of T-line antigen streak solution: 100 mM PB buffer containing 1.5 mg / mL of atrazine complete antigen (atrazine-ovalbumin conjugate), PEG20000 with a concentration of 0.5% by mass, pH The value is 6.2;

[0075] Prepare C-line antibody streak solution: containing 0.3mg / mL goat anti-mouse antibody, diluted with PBS solution.

[0076] The T-line antigen streak solution and the C-line antibody streak solution were respectively streaked and coated on the NC membrane. The streaking flow rate was 1 μL / cm, and the distance between the T line and the C line was 5 mm. Place the scratched NC film in an oven at 37°C for 4 hours and take it out.

[0077] The colloidal gold labeled with the atrazine monoclonal antibody prepared in Example 1 was lyophilized and stored on the reaction well, and the sample pad, binding pad, NC membrane, and water-absorbing pad were pasted.

[0078] Specific paste method: first paste the...

experiment example 1

[0083] Experimental example 1 Preparation and optimization of atrazine monoclonal antibody-colloidal gold marker

[0084] Take 1 mL of colloidal gold, add 20 μL of 0.1M sodium carbonate to adjust the pH to about 8.5, add 3, 5, 7, 10 μL of atrazine monoclonal antibody with a concentration of 1 mg / mL, mix for 30 minutes, add 120 μL of 10% BSA, and block for 15 minutes , centrifuge at 10000rpm for 15min, discard the supernatant, add 1mL of antibody stabilization solution, redissolve the precipitate, continue to centrifuge at 8000rpm for 15min, repeat twice, and finally resuspend in 100μL of antibody stabilization solution to obtain colloidal gold labeled with atrazine monoclonal antibody.

[0085] The prepared atrazine monoclonal antibody-labeled colloidal gold was mixed with 50ppb ATZ for incubation, and then added dropwise to the sample pad of the test strip, and a blank group (only containing atrazine monoclonal antibody-labeled colloid gold) as a control. The atrazine monocl...

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Abstract

The invention relates to the field of biological detection, in particular to a detection preparation for detecting atrazine, a kit thereof and a detection method. The detection preparation comprises a colloidal gold test strip and a reaction hole, the colloidal gold test strip comprises a sample pad, an NC membrane and an absorption pad, and a T line and a C line are arranged on the NC membrane; colloidal gold labeled by an atrazine monoclonal antibody is freeze-dried and stored in the reaction hole; the T line is coated with an atrazine complete antigen and is used for competitively combining the colloidal gold labeled by the atrazine monoclonal antibody; and the C line is coated with a goat anti-mouse antibody and is used for capturing uncombined colloidal gold labeled by the atrazine monoclonal antibody. The detection preparation provided by the invention can detect atrazine within 15 minutes, and has the technical advantages of wide linear range, low detection limit and high sensitivity.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a detection preparation for detecting atrazine, a kit and a detection method thereof. Background technique [0002] Atrazine (ATZ) is also known as atrazine, and its molecular formula is C 8 h 14 ClN 5 . It is a typical triazine herbicide. Atrazine is a selective systemic conduction pre-emergence and post-emergence herbicide. It is mainly used in dry fields such as corn, sorghum, sugarcane, fruit trees, and woodlands to control annual grasses. Weeds and broad-leaved weeds also have a certain inhibitory effect on certain perennial weeds. Although it is a low toxicity herbicide, it is moderately persistent in soil with a half-life of up to 57 weeks. The pollution caused by atrazine due to its stable chemical properties and long residual time has attracted more and more attention. Atrazine was positively detected in surface water, groundwater, soil, and crops. ATZ was found...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/558G01N33/58G01N33/543G01N33/53
CPCG01N33/577G01N33/558G01N33/587G01N33/54346G01N33/5308G01N2430/20
Inventor 孙铁强姚站馨宁保安高蔚娜张予弦郭长江杨涵王新阳
Owner INST OF ENVIRONMENTAL MEDICINE & OCCUPATIONAL MEDICINE ACAD OF MILITARY MEDICINE ACAD OF MILITARY SCI
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