Nitrofurans drug metabolite assay kit and production method thereof

A detection kit and a technology for nitrofurantoin metabolites are applied in the field of biological immunoassay determination, and can solve the problems of cumbersome, time-consuming, expensive instruments and the like for detection

Inactive Publication Date: 2012-10-31
宝瑞源生物技术(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods have high sensitivity and accurate results, but have the disadvantages of requiring expensive instruments, cumbersome and time-consuming detection, and the detection personnel need certain professional training.

Method used

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  • Nitrofurans drug metabolite assay kit and production method thereof
  • Nitrofurans drug metabolite assay kit and production method thereof
  • Nitrofurans drug metabolite assay kit and production method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Preparation of Nitrofuran Drug Metabolite Detection Kit

[0041] 1. Preparation of nitrofuran drug metabolite conjugated BSA

[0042] AOZ, AMOZ, SEM and AHD haptens were coupled with bovine serum albumin by active ester method to obtain AOZ-BSA conjugates, AMOZ-BSA conjugates, SEM-BSA conjugates and AHD-BSA conjugates As an immunogen and coater.

[0043] 2. Preparation of monoclonal antibodies to nitrofuran drug metabolites

[0044] AOZ-BSA conjugates were used as immunogens to immunize BALB / C mice to prepare AOZ monoclonal antibodies; AMOZ-BSA conjugates were used as immunogens to immunize BALB / C mice to prepare AMOZ monoclonal antibodies; SEM-BSA conjugates BALB / C mice were immunized as immunogen to prepare SEM monoclonal antibody; AHD-BSA conjugate was used as immunogen to immunize BALB / C mice to prepare AHD-monoclonal antibody. .

[0045] 3. Preparation of colloidal gold

[0046] Take 100mL of 0.01% chloroauric acid aqueous solution, heat to boil. Q...

Embodiment 2

[0062] Example 2: Detection of nitrofuran drug metabolite detection kit

[0063] 1. Sample handling

[0064] After taking 1g of tissue sample and crushing the tissue sample, add 5ml of distilled water, 0.5ml of 1M HCl and 100μl of 0.01M benzaldehyde (ethanol solution), shake fully, and place at 38°C overnight. Then add 0.1M of K 2 HPO 4 5ml, 0.4ml of 1M NaOH and 5ml of ethyl acetate, shake vigorously for 30 seconds. Centrifuge at 4000 rpm for 10 minutes at room temperature, then transfer 2.5 ml of the ethyl acetate supernatant to a new container. It was blown dry with nitrogen at 45° C., redissolved in n-hexane and mixed with 1 ml of 0.01 M PBST (pH 7.4, 0.05% Tween-20). Centrifuge at 4000 rpm for 10 minutes at room temperature, and finally take the bottom aqueous phase for determination.

[0065] 2. Detection method:

[0066] Take out the nitrofuran drug metabolite detection kit and place it horizontally; drop 3 drops of sample on the sample pad, observe and record the...

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Abstract

The invention provides a nitrofurans drug metabolite assay kit, which consists of a furazolidone metabolite test paper strip, a furaltadone metabolite test paper strip, a nitrofurazone metabolite test paper strip and a furantoin metabolite test paper strip. Each test paper strip consists of a sample pad, a colloidal gold pad, a nitrocellulose membrane, a sample adsorbing pad and a PVC (polyvinyl chloride) supporting plate, wherein the colloidal gold layer is a nitrofurans drug metabolite monoclonal antibody polyester film marked with colloidal gold, the nitrocellulose membrane is sequentially coated with nitrofurans drug metabolite coupled carrier protein as a test line (a T-line) and goat anti-mouse IgG antibody as a quality control line (a C-line). The nitrofurans drug metabolite assay kit is produced according to the nano colloidal gold technology, antigen-antibody specificity reactions and principles of immunity competitive inhibition reaction, and is used for testing whether samples contain nitrofurans drug metabolites or not.

Description

technical field [0001] The invention relates to the technical field of determination of biological immunological methods, in particular to a detection kit for metabolites of nitrofuran drugs produced by colloidal gold immune layer technology and a preparation method thereof. Background technique [0002] Nitrofuran drugs are a kind of spectrum antibiotics, mainly referring to furaltadone, furazolidone, nitrofurantoin, and nitrofurazone. They have good antibacterial effects and are widely used in feed additives and therapeutic drugs. Gastrointestinal diseases of pigs, cattle, poultry and bees caused by Helicobacter. They have adverse effects on human health and are carcinogenic. For the sake of food safety, the European Union listed nitrofuran antibiotics as Class A prohibited drugs in the 96 / 23 / EC regulation, and decided to ban the addition of any nitrofuran in animal feed from January 1, 1997 antibiotic. my country also issued a ban on the use of nitrofuran antibiotics i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/532
Inventor 陈立柱杨利李峰
Owner 宝瑞源生物技术(北京)有限公司
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