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Magnetic particle chemiluminescence kit for detecting nitrofurantion and application thereof

A chemiluminescence reagent, nitrofurantoin technology, applied in chemiluminescence/bioluminescence, analysis through chemical reaction of materials, measurement devices, etc., can solve problems such as poor stability of reagents, reaction time and temperature influence, and achieve detection time Low, fast detection, high sensitivity effect

Inactive Publication Date: 2012-07-04
BEIJING KWINBON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The reaction time and temperature have a great influence on the enzyme activity, and the stability of the reagent is poor

Method used

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  • Magnetic particle chemiluminescence kit for detecting nitrofurantion and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The preparation of embodiment 1 kit specific components

[0036] 1. Preparation of luminescent markers

[0037] a) Synthesis of haptens

[0038] Nitrofurantoin and p-carboxybenzaldehyde are derivatized by reacting in water to obtain a nitrofurantoin derivative hapten.

[0039] Dissolve 500 mg of nitrofurantoin in 5 ml of pure water. Dissolve 500 mg of p-carboxybenzaldehyde in 15 ml of water, add it to the nitrofurantoin solution and react at room temperature for 48 hours to obtain a light yellow precipitate which is the nitrofurantoin derivative hapten. The nitrofurantoin derivative hapten is obtained by washing repeatedly 5-6 times with 25 ml of water and drying.

[0040] b) Preparation of luminescent markers

[0041] Take 4.5mmol / L ABEI, dissolve it in 4ml distilled water, dissolve 5.0mmol / L N-hydroxysuccinimide in 0.5ml N,N-dimethylformamide, mix the two thoroughly and react at room temperature for 3-4h . Take 15 mg of the above-prepared nitrofurantoin hapten, ...

Embodiment 2

[0057] The formation of embodiment two kits

[0058] The magnetic particle chemiluminescent detection kit for detecting nitrofurantoin was assembled so that it contained the following components:

[0059] Fluorescent marker of FITC-labeled nitrofurantoin monoclonal antibody

[0060] Luminescent marker of ABEI-labeled nitrofurantoin hapten

[0061] Separation reagent of paramagnetic nanobeads coated with goat anti-FITC monoclonal antibody

[0062] Nitrofurantoin standard solution (0ng / ml, 0.01ng / ml, 0.03ng / ml, 0.09ng / ml, 0.27ng / ml, 0.81ng / ml), the standard dilution is pH7.4, containing 0.05% thimerosal preservative , 0.1mol / LTRIS buffer. The percentage content is a mass percentage content.

[0063] The concentrations of the nitrofurantoin quality control solution were 0.02ng / ml and 0.5ng / ml respectively, and the quality control substance was diluted to pH 7.4, containing 0.05% thimerosal preservative, and 0.1mol / LTRIS buffer. The percentage content is a mass percentage con...

Embodiment 3

[0065] The detection of nitrofurantoin in the actual sample of embodiment three

[0066] 1. Pretreatment of feed samples

[0067] Weigh 1.0g±0.05g feed sample into a 50ml centrifuge tube, add 8ml ethyl acetate, then add 2ml acetonitrile; shake for 5 minutes, rest for 10 minutes, room temperature above 3000g, centrifuge for 10 minutes, take 2ml of supernatant to 10ml of clean glass In the test tube, blow dry under nitrogen flow in a water bath at 50-60°C; add 1ml of n-hexane, vortex for 1min with a vortexer, then add 1ml of reconstituted working solution, and vortex for 30s with a vortexer; over 3000g, room temperature (20- Centrifuge at 25°C for 10 min; remove the upper organic phase, take 200 μl of the lower aqueous phase and mix with 200 μl of reconstitution working solution; take 50 μl for analysis, sample dilution factor: 10.

[0068] 2. Detection with kit and analysis of results

[0069] Pipette 20μl-100μl standard or sample respectively, then add 20μl-100μl luminescent...

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Abstract

The invention relates to a magnetic particle chemiluminescence kit for detecting nitrofurantion. The kit comprises the following reagents: a luminescent label, a fluorescent label, a standard product, a quality control product and a separating reagent. The luminescent label is of a nitrofurantion hapten labeled by an isoluminol luminescent label, the fluorescent label is of a nitrofurantion monoclonal antibody labeled by fluorescein or derivatives thereof, and the separating reagent is of paramagnetic nano micro-beads coated with a goat anti-FITC (fluorescein isothiocyanate) monoclonal antibody. The invention further relates to a method for detecting the nitrofurantion in feed by adopting the kit, and the method has the advantages of higher sensitivity in detection of the nitrofurantion, higher specificity and faster detection speed.

Description

technical field [0001] The invention relates to a direct chemiluminescence detection kit and a testing method thereof, in particular to a magnetic particle competition direct chemiluminescence detection kit for detecting nitrofurantoin residues in feed samples. technical background [0002] Nitrofuran drugs are synthetic broad-spectrum antibiotics, which have very good antibacterial effects and pharmacokinetic properties, and have been widely used as growth-promoting additives for livestock, poultry, and artificially farmed aquatic products. However, long-term experimental studies have found that both nitrofuran drugs and metabolites can cause cancer and gene mutations in experimental animals, and it can be inferred that humans have long-term use of such drugs or long-term consumption of poultry, Livestock and artificially raised aquatic products can also undergo canceration and gene mutations. Therefore, such drugs are prohibited from being used in treatment and feed. [...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76G01N33/577G01N33/533
Inventor 何方洋万宇平罗晓琴常平平陈炜玲蒲小蓉段盈盈
Owner BEIJING KWINBON BIOTECH
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