A kind of nitrofurantoin chemiluminescent detection kit

Abstract

The invention relates to a chemiluminescence detection kit used for detecting macrodantin. The chemiluminescence detection kit comprises a wash concentrate, a negative reference substance, a positive reference substance, a chemiluminescence liquid, macrodantin standard substance, horse radish peroxidase labelled macrodantin specific antibody, and a luminescence test plate coated with macrodantin. The horse radish peroxidase labelled macrodantin specific antibody is horse radish peroxidase labelled anti-macrodantin polyclonal antibody obtained via immunization of New Zealand white rabbits, wherein macrodantin-BSA is taken as an immunogen; and the horse radish peroxidase labelled macrodantin specific antibody is prepared via adoption of sodium periodate method. The chemiluminescence detection kit used for detecting macrodantin is high in sensitivity, and wide in detection range; is simple and convenient for operation; and possesses no toxicity. No pollution is caused by the chemiluminescence detection kit. According to the chemiluminescence detection kit, the envelope antigen is prepared via coupling of a macrodantin metabolite and a carrier, so that cost is low. The horse radish peroxidase labelled macrodantin specific antibody is prepared via one-step method, so that the preparation method and preparation steps are simpler, and it is beneficial for industrialized production.

Description

technical field

[0001] The invention relates to a kit, in particular to a nitrofurantoin chemiluminescence detection kit. Background technique

[0002] At present, the techniques for detecting antibiotics in feed mainly include the following methods:

[0003] (1) Physical and chemical detection method

[0004] After the 1990s, the vast majority of physical and chemical detection methods for the determination of nitrofurantoin mainly rely on liquid chromatography for separation, followed by chromatography / mass spectrometry, gas chromatography, and high-performance thin-layer chromatography. It has a slight application in the detection of antibiotic residues. Chromatographic detection of drug residues in feed requires sample processing (including sample extraction, deproteinization, centrifugation, chromatographic column purification, derivatization, etc.), drug separation and drug detection. The physical and chemical detection method uses the special reaction or property o...

Examples

preparation example Construction

[0056] Step 3, preparation of horseradish peroxidase marker

[0057] The enzyme-labeled nitrofurantoin antibody was prepared by the sodium periodate method, and the reaction system was as follows: first use 1ml of horseradish peroxidase solution with a concentration of 5mg / ml and 0.15ml of NaIO with a concentration of 0.1mol / L 4 After mixing the solution, place it at 4°C in the dark for 0.5 hours, then add 0.25ml of ethylene glycol and shake it well at room temperature in the dark for 1 hour, then place it at 4°C for dialysis overnight, and then use 0.2ml of 0.2mol / L with a pH value of Adjust the pH value of the hydroformylated horseradish peroxidase solution obtained after dialysis to be 9.2-9.5 in the carbonate coating solution of 9.6, dissolve 5 mg of anti-nitrofurantoin polyclonal antibody in 1.0 ml concentration of 0.01mol / L, pH9. Immediately after the coating solution of 6, add the above-mentioned horseradish peroxidase solution after hydroformylation and react at 37°C f...

Embodiment

[0074] A nitrofurantoin chemiluminescence detection kit, including concentrated lotion, negative control substance, positive control substance, chemiluminescent solution, nitrofurantoin standard substance, horseradish peroxidase-labeled nitrofurantoin-specific antibody and nitrofurantoin-coated luminescence plate ;

[0075] The concentrated lotion is: phosphate buffer containing Tween-20 and sodium chloride;

[0076] The negative control substance is: more than 5 parts of the pig urine mixture that is negative for nitrofurantoin after sterilization and filtration stored separately;

[0077] The positive control substance is: more than 5 parts of pig urine mixture that is positive for nitrofurantoin after sterilization and filtration stored separately;

[0078]The chemiluminescent solution includes liquid A and liquid B; the preparation method of liquid A is as follows: first configure a Tris-HCl buffer solution of 0.2mol / LpH7.8, add luminol and sodium tetraphenylborate to thi...

technical field

[0001] The invention relates to a kit, in particular to a nitrofurantoin chemiluminescence detection kit. Background technique

[0002] At present, the techniques for detecting antibiotics in feed mainly include the following methods:

[0003] (1) Physical and chemical detection method

[0004] After the 1990s, the vast majority of physical and chemical detection methods for the determination of nitrofurantoin mainly rely on liquid chromatography for separation, followed by chromatography / mass spectrometry, gas chromatography, and high-performance thin-layer chromatography. It has a slight application in the detection of antibiotic residues. Chromatographic detection of drug residues in feed requires sample processing (including sample extraction, deproteinization, centrifugation, chromatographic column purification, derivatization, etc.), drug separation and drug detection. The physical and chemical detection method uses the special reaction or property o...

Examples

preparation example Construction

[0056] Step 3, preparation of horseradish peroxidase marker

[0057] The enzyme-labeled nitrofurantoin antibody was prepared by the sodium periodate method, and the reaction system was as follows: first use 1ml of horseradish peroxidase solution with a concentration of 5mg / ml and 0.15ml of NaIO with a concentration of 0.1mol / L 4 After mixing the solution, place it at 4°C in the dark for 0.5 hours, then add 0.25ml of ethylene glycol and shake it well at room temperature in the dark for 1 hour, then place it at 4°C for dialysis overnight, and then use 0.2ml of 0.2mol / L with a pH value of Adjust the pH value of the hydroformylated horseradish peroxidase solution obtained after dialysis to be 9.2-9.5 in the carbonate coating solution of 9.6, dissolve 5 mg of anti-nitrofurantoin polyclonal antibody in 1.0 ml concentration of 0.01mol / L, pH9. Immediately after the coating solution of 6, add the above-mentioned horseradish peroxidase solution after hydroformylation and react at 37°C f...

Embodiment

[0074] A nitrofurantoin chemiluminescence detection kit, including concentrated lotion, negative control substance, positive control substance, chemiluminescent solution, nitrofurantoin standard substance, horseradish peroxidase-labeled nitrofurantoin-specific antibody and nitrofurantoin-coated luminescence plate ;

[0075] The concentrated lotion is: phosphate buffer containing Tween-20 and sodium chloride;

[0076] The negative control substance is: more than 5 parts of the pig urine mixture that is negative for nitrofurantoin after sterilization and filtration stored separately;

[0077] The positive control substance is: more than 5 parts of pig urine mixture that is positive for nitrofurantoin after sterilization and filtration stored separately;

[0078]The chemiluminescent solution includes liquid A and liquid B; the preparation method of liquid A is as follows: first configure a Tris-HCl buffer solution of 0.2mol / LpH7.8, add luminol and sodium tetraphenylborate to thi...

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