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Chemiluminescent immunoassay kit for detecting o-phenylphenol in fruits and vegetables

A chemiluminescent immunoassay and o-phenylphenol technology, applied in the field of immunological detection, can solve the problems of toxicity, damage to liver and kidney, nervous system and bone marrow, etc., and achieve the effect of simple instrument, increased sensitivity and low detection cost

Inactive Publication Date: 2019-05-14
JIANGSU WISE SCI & TECH DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the residual preservatives in the fruit have certain toxicity to the human body, mainly infringing on the liver, kidney, nervous system and bone marrow of the human body

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] 1. Preparation of each component of the kit

[0040] (1) Preparation of o-phenylphenol hapten: o-phenylphenol was acidified, and reacted with sodium nitrite in a low-temperature environment without light at 4°C to generate an intermediate containing diazonium cations. The diazotized o-phenylphenol is used as a hapten for subsequent synthesis of immune antigens and coating antigens.

[0041] (2) Preparation of o-phenylphenol-bovine serum albumin (BSA) immunogen: o-phenylphenol and bovine serum albumin (BSA) were coupled by a diazotization method to obtain an immune antigen.

[0042] Preparation of o-phenylphenol-oval serum albumin (OVA) coated antigen: o-phenylphenol and oval serum albumin (OVA) were coupled by diazotization method to obtain the coated antigen.

[0043]Preparation of o-phenylphenol-peroxidase-labeled antibody: For 6-8 week-old female BALB / c mice (body weight 18-20 g), the high-dose immunization protocol is as follows: 160 µg o-phenylphenol is used for t...

Embodiment 2

[0072] A chemiluminescent immunoassay kit for the detection of o-phenylphenol, comprising the following components:

[0073] (1) 96-well opaque white ELISA plate (8 wells x 12 strips) is coated with o-phenylphenol-mouse serum albumin conjugate, and is vacuum-sealed in an aluminum foil bag.

[0074] (2) 6 bottles of o-phenylphenol standard solution, the concentrations are:

[0075] 0 ng / mL, 0.02 ng / mL, 0.08 ng / mL, 0.32 ng / mL, 1.28 ng / mL, 5.12 ng / mL

[0076] (3) Ortho-phenylphenol-horseradish peroxidase labeled antibody solution.

[0077] (4) Luminescence substrate A solution (luminol and enhancer), luminescence substrate B solution (urea hydrogen peroxide).

[0078] (5) 2 times concentrated sample diluent. Before use, please dilute 1:1 (1 part concentrated diluent + 1 part deionized water) to become the working sample diluent, and the diluted working sample diluent is 0.05 mol / L, pH 7.4 Tris-HCl buffer.

[0079] (6) 20 times concentrated washing solution. Before use, pleas...

Embodiment 3

[0081] A chemiluminescent immunoassay kit for the detection of o-phenylphenol, comprising the following components:

[0082] (1) A 96-well opaque white microplate (8 wells x 12 strips) is coated with o-phenylphenol-egg albumin conjugate, and is vacuum-sealed in an aluminum foil bag.

[0083] (2) 6 bottles of o-phenylphenol standard solution, the concentrations are:

[0084] 0 ng / mL, 0.02 ng / mL, 0.08 ng / mL, 0.32 ng / mL, 1.28 ng / mL, 5.12 ng / mL

[0085] (3) Ortho-phenylphenol-horseradish peroxidase labeled antibody solution.

[0086] (4) Luminescence substrate A solution (luminol and enhancer), luminescence substrate B solution (urea hydrogen peroxide).

[0087] (5) 2 times concentrated sample diluent. Before use, please dilute 1:1 (1 part of concentrated diluent + 1 part of deionized water) to become the working sample diluent, and the diluted working sample diluent is 0.05 mol / L, pH 7.4 glycine-HCl buffer.

[0088] (6) 20 times concentrated washing solution. Before use, ple...

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PUM

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Abstract

The invention provides a chemiluminescent immunoassay kit for detecting o-phenylphenol, and belongs to the field of immunological detection. The kit consists of an opaque white ELISA (Enzyme Linked Immunosorbent Assay) plate coated with an o-phenylphenol-carrier protein conjugate, an o-phenylphenol standard, an o-phenylphenol-peroxidase labelled antibody working fluid, a luminous substrate solution, a concentrated sample diluent and a concentrated washing liquid, wherein the o-phenylphenol-carrier protein conjugate is obtained by coupling o-phenylphenol with carrier proteins by a mixed anhydride method or a carbodiimide method, and the concentrated washing liquid contains 0.05 percent of Tween-20. Compared with traditional enzyme-linked immunosorbent assay, the kit provided by the invention has the advantages of higher sensitivity, shorter detection time and lower cost, and can be applied to the residual amount detection of o-phenylphenol in apple samples.

Description

technical field [0001] The invention relates to a chemiluminescent immunoassay kit for detecting o-phenylphenol, which is used for detecting the content or residual amount of o-phenylphenol in fruits and vegetables in animal-derived foods. It belongs to the field of immunological detection. Background technique [0002] Food safety and hygiene are related to human life and health, and have aroused widespread concern from the international community. With the reduction of natural ways of food production and the increase of the proportion of industrialization, the chances of food being intentionally or unintentionally contaminated by people are gradually increasing. The size is also unprecedented. The current fresh-keeping storage technology of fruits and vegetables mainly relies on suitable low temperature and air conditioning, but the growth of mold cannot be controlled under the temperature and gas conditions that they can withstand during the storage of fruits and vegeta...

Claims

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Application Information

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IPC IPC(8): G01N33/535G01N33/577
Inventor 杜霞洪霞丁炎
Owner JIANGSU WISE SCI & TECH DEV
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