Preparation method of chemiluminescence immunoassay detection kit used for detecting methyl parathion

A technique of chemiluminescence immunity and methyl parathion, which is applied in the field of immunological detection, can solve the problems of difficult to meet the on-site, batch, rapid detection of samples, high detection cost, complicated processing, etc., and achieves low detection cost and increased sensitivity. , the effect of a wide linear range

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

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] 1. Preparation of each component of the kit

[0039] (1) Preparation of methyl parathion hapten: acidify methyl parathion (MP) and react with sodium nitrite in a low temperature environment with no light at 4°C to generate an intermediate containing diazonium cations. Diazotized methyl parathion (MP) is used as a hapten to synthesize immune antigens and coating antigens;

[0040] (2) Preparation of methyl parathion-bovine serum albumin (BSA) immunogen: the methyl parathion and bovine serum albumin (BSA) are coupled by diazotization method to obtain immune antigen.

[0041] Preparation of methyl parathion-ovum serum albumin (OVA) coating antigen: the methyl parathion and oval serum albumin (OVA) are coupled by diazotization method to obtain the coating antigen.

[0042] Preparation of methyl parathion-peroxidase-labeled antibody: For female BALB / c mice aged 6-8 weeks (body weight 18-20g), the high-dose immunization protocol is 160μg MP-BSA and the same amount for the first immun...

Embodiment 2

[0068] The chemiluminescence immunoassay kit for detecting methyl parathion includes the following components:

[0069] (1) 96-well opaque white ELISA plate (8-well × 12 strips) is coated with methyl parathion-OVA conjugate, and vacuum-sealed packaging with aluminum foil bag;

[0070] (2) 6 bottles of methyl parathion standard solution, the concentrations are:

[0071] 0ng / mL, 0.01ng / mL, 0.05ng / mL, 0.5ng / mL, 5ng / mL, 25ng / mL

[0072] (3) Methyl parathion-horseradish peroxidase labeled antibody solution;

[0073] (4) Luminescent substrate A liquid (luminol and enhancer), luminescent substrate B liquid (urea hydrogen peroxide);

[0074] (5) Concentrate the sample diluent twice. Before use, please dilute 1:1 (1 part concentrated diluent + 1 part deionized water) to become the working sample diluent. The diluted working sample diluent is 0.05mol / L, pH7.4 Tris-HCl buffer ;

[0075] (6) Concentrate the washing solution 20 times. Before use, please dilute 1:19 (1 part concentrated diluent + 19...

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Abstract

The invention provides a chemiluminescence immunoassay detection kit used for detecting methyl parathion (MP), and belongs to the field of immunological detection. The chemiluminescence immunoassay detection kit is composed of a non-transparent white enzyme label plate coated with a methyl parathion (MP)-carrier protein conjugate, a methyl parathion standard substance, a methyl parathion-peroxidase labelled antibody working liquid, a luminescent substrate liquid, a concentrated sample diluent, and a concentrated washing liquid. The methyl parathion (MP)-carrier protein conjugate is obtained via coupling of methyl parathion with a carrier protein via mixed anhydride method or carbodiimide method; and the concentrated washing liquid contains 0.05% of tween-20. The chemiluminescence immunoassay detection kit is high in sensitivity; detection time is short; cost is low; and the chemiluminescence immunoassay detection kit can be used for residual amount detection of methyl parathion (MP) in samples such as fruits, vegetables, and crops.

Description

Technical field [0001] The invention relates to a chemiluminescence immunoassay kit for detecting methyl parathion (MP), which is used for detecting the content or residual amount of methyl parathion (MP) in fruits, vegetables, crops and the like. It belongs to the field of immunological detection. Background technique [0002] Parathionmethyl (Parathionmethyl), also called O,O-dimethyl-O-p-nitrophenyl thiophosphate, is a highly toxic organophosphate insecticide with large output and wide application. It is highly toxic and has been banned on vegetables and fruits in recent years. However, in vegetable production, there are still the phenomenon of misuse and abuse of pesticides. In the supervision and testing of import and export trade of agricultural products and food safety, methyl parathion (MP) is still the focus of routine testing. [0003] Methyl parathion (MP) residue analysis generally uses gas chromatography (GC), high performance liquid chromatography (HPLC) and gas chr...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/577
Inventor 洪霞杜霞
Owner JIANGSU WISE SCI & TECH DEV
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