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Preparation of chemiluminescence immunoassay kit for detecting chloroquine

A chemiluminescent immunoassay and detection kit technology, which is applied in the field of immunological detection, can solve the problems that it is difficult to meet the sample site, batch, rapid detection, high detection cost, tedious and time-consuming, etc., and achieve low detection cost, increased sensitivity, and reduced The effect of using links

Inactive Publication Date: 2015-05-27
NANJING YITE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Chloroquine 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 drugs, but require expensive Instruments, sample pretreatment is complex, tedious and time-consuming, the detection cost is high, and requires professional operation, it is difficult to meet the needs of on-site, batch, and rapid detection of samples

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 chloroquine hapten: acidify chloroquine, and react with sodium nitrite in a low-temperature environment without light at 4°C to generate an intermediate containing diazonium cations. The diazotized chloroquine is used as a hapten for subsequent synthesis of immune antigens and coating antigens;

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

[0041] Preparation of chloroquine-oval serum albumin (OVA) coated antigen: chloroquine and egg serum albumin (OVA) were coupled by diazotization method to obtain coated antigen.

[0042] Preparation of chloroquine-peroxidase-labeled antibody: For 6-8 week-old female BALB / c mice (body weight 18-20 g), the high-dose immunization scheme is as follows: 160 μg chloroquine-BSA and an equal amount of Freud for the first immunization...

Embodiment 2

[0071] The chemiluminescent immunoassay kit for detecting chloroquine includes the following components:

[0072] (1) 96-well opaque white ELISA plate (8 wells x 12 strips) coated with chloroquine-OVA conjugates, vacuum-sealed in aluminum foil bags;

[0073] (2) 6 bottles of chloroquine standard solution, the concentrations are respectively:

[0074] 0 ng / mL, 0.01 ng / mL, 0.05 ng / mL, 0.5 ng / mL, 5 ng / mL, 25 ng / mL

[0075] (3) chloroquine-horseradish peroxidase labeled antibody solution;

[0076] (4) Luminescence substrate A solution (luminol and enhancer), luminescence substrate B solution (urea hydrogen peroxide);

[0077] (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 Tris-HCl buffer;

[0078] (6) 20 times concentrated washing solution. Before use, please dilute at 1:19 (1 part of concentrat...

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Abstract

The invention provides a preparation of a chemiluminescence immunoassay kit for detecting chloroquine and belongs to the field of immunological detection. The kit is composed of: a non-transparent white enzyme label plate coated by a chloroquine-carrier protein conjugate, a chloroquine standard substance, a chloroquine-peroxidase marked antibody operating fluid, a luminescent substrate liquid, a concentrated sample diluting liquid and a concentrated washing liquid. The chloroquine-carrier protein conjugate is prepared by coupling chloroquine to a carrier protein in a mixed anhydride method or a carbodiimide method. The concentrated washing liquid contains 0.05% of tween-20. The kit is higher in sensitivity, is short in detection time, is low in cost and can be used for detection of residual amount of the chloroquine in samples such as fruits, crops and vegetables and the like.

Description

technical field [0001] The invention relates to a chemiluminescent immunoassay kit for detecting chloroquine, which is used for detecting the content or residual amount of chloroquine in fruits, vegetables, crops and the like. It belongs to the field of immunological detection. Background technique [0002] Chloroquine metabolizes slowly in the human body, and when the content exceeds a certain amount, it is easy to cause reactions such as loss of appetite, nausea and vomiting, and diarrhea; skin itching, purpura, hair loss, whitening of hair, eczema, exfoliative dermatitis, and psoriasis can also occur; heavy head, Headache, dizziness, tinnitus, vertigo, fatigue, sleep disturbance, mental confusion, reduced vision, corneal and retinal degeneration, etc. [0003] Chloroquine 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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/543
CPCG01N33/54393G01N21/763G01N2430/00
Inventor 洪霞其他发明人请求不公开姓名
Owner NANJING YITE BIOTECH
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