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Time-resolved fluorescence immunoassay kit detecting methyl parathion and detection method thereof

A technology of time-resolved fluorescence and methyl parathion, which is applied in the direction of measuring devices, analytical materials, instruments, etc., can solve the problems that it is difficult to meet the requirements of sample site, batch, rapid detection, high detection cost, cumbersome and time-consuming, etc., and achieve the goal of operation Simple, widely used, and highly specific effects

Inactive Publication Date: 2016-02-10
JIANGSU WISE SCI & TECH DEV
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Problems solved by technology

[0003] Methyl parathion (MP) residue analysis generally uses gas chromatography (GC), high performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC / MS). These methods are sensitive and accurate, and can simultaneously determine multiple It is a kind of drug, but it needs expensive instruments, the sample pretreatment is complex, tedious and time-consuming, the detection cost is high, and it needs to be operated by professionals, it is difficult to meet the needs of on-site, batch and rapid detection of samples
There are currently no patents and literature reports on a chronofluoroimmunoassay for the detection of methylparathion (MP)

Method used

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  • Time-resolved fluorescence immunoassay kit detecting methyl parathion and detection method thereof

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Embodiment

[0023] 1. Preparation of immunogen and coating agent

[0024] Synthesis of the immunogen (MP-BSA) of the present invention: Accurately weigh 324 mg of methyl parathion and dissolve it in 2 mL of N,N-dimethylformamide, add γ-aminobutyric acid solution drop by drop under stirring, and stir for 3 hours , adjust the pH of the reaction solution to about 10. The precipitate was removed by centrifugation. Add the above reaction dropwise to BSA solution (320mgBSA dissolved in 5mL normal saline), then add 23mg of N-hydroxysuccinimide (NHS), 45.4mg of N,N-dicyclohexylcarbodiimide (DCC), 4 React overnight at ℃, centrifuge to remove the precipitate, take the supernatant and dialyze with phosphate buffered solution (PBS) for 3 days, change the dialysate every 6 hours, freeze-dry the obtained product, and store it at -20℃ for later use;

[0025] Synthesis of the coating agent (MP-OVA): In the above reaction, after replacing BSA with OVA, the reaction conjugate MP-OVA was obtained, which w...

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Abstract

The invention discloses a time-resolved fluorescence immunoassay kit detecting methyl parathion (MP) and a detection method thereof. The time-resolved fluorescence immunoassay kit detecting methyl parathion is composed of a porous coating plate, a buffer, a methyl parathion standard sample, a methyl parathion antibody freeze-dried sample, an europium-labelled goat anti-mouse antibody, a washing liquid and an enhancing liquid. The detection method employing the time-resolved fluorescence immunoassay kit detecting methyl parathion comprises the following steps: (1) preparing an immunogen; (2) preparing a coating antigen; (3) preparing a monoclonal antibody; and performing pretreatment and detection on a sample. The kit and the method are short in detection time, high in average recovery rate, simple for sample pretreatment, capable of realizing on-site operation detection, wide in application scope and low in detection cost, and also possess the advantages of being strong in detection specificity, small in intra- and inter-assay difference, high in sensitivity, simple and rapid in operation, especially suitable for detection of a large bath of samples, and the like.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a time-resolved fluorescent immunoassay kit for detecting methyl parathion (MP) and a detection method thereof. Background technique [0002] Parathionmethyl, also known as O,O-dimethyl-O-p-nitrophenylphosphorothioate, is a highly toxic organophosphate insecticide with large output and wide application. It is highly toxic and has been banned from use on vegetables and fruits in recent years. However, indiscriminate use and abuse of pesticides still exist in vegetable production, and methyl parathion (MP) is still the focus of routine testing in agricultural product import and export trade and food safety supervision and testing. [0003] Methyl parathion (MP) residue analysis generally uses gas chromatography (GC), high performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC / MS). These methods are sensitive and accurate, and can simul...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
Inventor 洪霞杜霞
Owner JIANGSU WISE SCI & TECH DEV
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