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System and method for preparing pesticide and veterinary drug residue multi-target nano enzyme probe and visually and rapidly detecting pesticide and veterinary drug residue multi-target nano enzyme probe

A detection method and detection system technology, applied in chemical instruments and methods, biological testing, physical/chemical process catalysts, etc., can solve the problems of unable to quickly screen samples, high cost, low sample detection efficiency, etc., and achieve visual rapid detection , low cost effect

Pending Publication Date: 2022-08-05
CHINA JILIANG UNIV
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Problems solved by technology

[0002] The quality and safety of agricultural products is a major issue related to people's livelihood. It requires not only social supervision but also the support of high-quality detection technology. There are many kinds of agricultural and veterinary drugs, and the routine detection technology process is cumbersome and costly. The current rapid detection technology is mostly aimed at a single target. The detection efficiency of samples coexisting with various hazard factors is low, and it is impossible to quickly screen a large number of samples on the spot. Therefore, it is particularly important to carry out rapid detection of agricultural and veterinary drug residues in edible agricultural products and develop related rapid detection products.

Method used

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  • System and method for preparing pesticide and veterinary drug residue multi-target nano enzyme probe and visually and rapidly detecting pesticide and veterinary drug residue multi-target nano enzyme probe
  • System and method for preparing pesticide and veterinary drug residue multi-target nano enzyme probe and visually and rapidly detecting pesticide and veterinary drug residue multi-target nano enzyme probe
  • System and method for preparing pesticide and veterinary drug residue multi-target nano enzyme probe and visually and rapidly detecting pesticide and veterinary drug residue multi-target nano enzyme probe

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Synthesis and identification of nanoenzyme Pt@Au materials and determination of catalytic function

[0036] Molarity of K at 20 mM 2 PtCl 4 and HAuCl 4 Solution, the mixing ratio is 9mL: 1mL, after mixing, add 5.5mL, 20mM ascorbic acid, add 0.8mL, 1mM Pluronic F127, the mixture is ultrasonically reacted in a water bath at room temperature for 3h, the product is separated by centrifugation at 10000r / min for 20min, and ethanol is mixed with ethanol. The residual Pluronic F127 was removed by repeated washing with water three times, and after reconstitution in ultrapure water, the concentration of Pt@Au nanomaterials was 12-20 mM, and the concentration was calculated as Pt. TEM samples were prepared for morphology and element distribution characterization, and the characterization results were as follows figure 1 As shown, the particle size of the synthesized Pt@Au nanozyme was 19.0±5.6 nm by statistical analysis, and the distribution analysis of Au and Pt elements found...

Embodiment 2

[0041] Preparation of Immune Signal Probes and Capture Probes for Various Pesticide and Veterinary Drug Residues

[0042] The preparation method of Pt@Au@Ab, a variety of pesticide and veterinary drug immune signal probes, includes: taking 4 centrifuge tubes, adding 3.0 mL of the above-mentioned Pt@Au nanomaterials diluted 10 times respectively, using 0.2M K 2 CO 3 Adjust the pH of the Pt@Au nanomaterial to 8.7, and add 20 μg chloramphenicol antibody, 40 μg carbendazim antibody, 50 μg omethoate antibody, and 30 μg salbutamol antibody in turn. After 1 h of reaction, add 30 μL PBS (0.01M, pH 7.4, containing 10 %BSA), react for 30 min to block the excess sites of Pt@Au, and after the incubation reaction, centrifuge at 10,000 r / min for 20 min to remove free antibody or protein to obtain the corresponding immune signal probe Pt@Au@Ab, which is stored at 4°C for later use.

[0043] Preparation of capture probes for various agricultural and veterinary drugs. Take 4 centrifuge tubes...

Embodiment 3

[0048] The feasibility of a rapid detection method based on immune signaling probes and capture probes was determined. Take 8 centrifuge tubes and add different pesticide and veterinary drug standards, capture probes and signal probes in sequence. After incubation for 30 min, remove the supernatant by magnetic separation. After washing the precipitate for 3 times, add 500.0 μL of complex substrate chromogenic solution in turn. , the B value of the reaction solution was read by a smartphone, as shown in Table 3, when no standard was added, the MPMs@Ag-Pt@Au@Ab immune complex was obtained by magnetic separation, and the signal probe with the most binding probe was detected. The maximum B value is obtained. When 50 μL of 10 μg / L corresponding standard is added, the analyte competes with the capture probe to bind the signal probe, and a small amount of signal probe is obtained by magnetic separation. The average detection B value of the system is compared with that without adding t...

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Abstract

The invention relates to a pesticide and veterinary drug residue multi-target nano enzyme immune probe preparation and visual rapid detection system and method, and the method comprises the following steps: preparing a Pt (at) Au nano material with a peroxidase catalysis function, preparing a signal probe Pt (at) Au (at) Ab corresponding to a target to be detected, and preparing a capture probe MPMs (at) Ag corresponding to the target to be detected. The rapid visual detection method is constructed by using a target nano-enzyme immunoprobe, and by optimizing test parameters and combining a magnetic separation technology, compared with a traditional enzyme-linked immunoassay method, the rapid visual detection method is more time-saving and rapid, and visual rapid detection of various pesticide and veterinary drug residues can be realized without the help of large instruments; the kit is applied to the visual rapid quantitative detection of carbendazim, omethoate, chloramphenicol and salbutamol, the detection limits are 0.01 mu g / L, 0.01 mu g / L, 0.05 mu g / L and 0.05 mu g / L in sequence, and the detection effect is excellent.

Description

technical field [0001] The invention belongs to the field of pesticide and veterinary drug residue detection, and relates to a nano-enzyme probe detection technology, in particular to a preparation and visual rapid detection system and method for a multi-target nano-enzyme probe of pesticide and veterinary drug residues. Background technique [0002] The quality and safety of agricultural products is a major issue related to people's livelihood. It requires not only social supervision but also the support of high-quality detection technology. There are many types of agricultural and veterinary drugs, and the routine detection technology process is cumbersome and expensive. The current rapid detection technology is mostly aimed at a single target. The detection efficiency of samples with coexistence of various hazard factors is low, and it is impossible to quickly screen a large number of samples on site. Therefore, it is particularly important to develop rapid detection produ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535G01N33/543G01N33/58G01N33/53B01J23/52
CPCG01N33/535G01N33/54326G01N33/581G01N33/5308B01J23/52Y02A40/70
Inventor 张彪张慧琪周若凡姚天择刘昀晖马宁申屠旭萍俞晓平
Owner CHINA JILIANG UNIV
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