Kit for quickly detecting content of chloroquine in crops
A kit and chloroquine technology, applied in the field of enzyme-linked immunosorbent assay kits for detecting chloroquine, can solve the problems of expensive equipment and complicated processing procedures, etc.
Inactive Publication Date: 2016-02-03
JIANGSU WISE SCI & TECH DEV
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[0003] At present, the determination methods of chloroquine mainly include high-performance liquid chromatography, reversed-phase high-performance liquid chromatography, surface-enhanced Raman spectroscopy, etc. The detection results are accurate and reliable, but the equipment is relatively expensive, and the sample pretreatment process is relatively complicated
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[0028] Preparation of Chloroquine Protein Conjugates:
[0029] Adopt the succinic anhydride method to obtain the chloroquine hapten derivative with carboxyl group, then get 0.05mmol and carrier protein BSA and mix in the carbonate buffer solution (CBS) of 0.05mol / LpH9.6 by the binding ratio of 10:1, then add 0.15mmol carbodiimide, stirred at room temperature for 24 hours, and finally dialyzed in 0.2mol / L PBS buffer solution of pH 7.6 for two days to remove unreacted haptens, and stored the obtained protein conjugate solution at -20°C spare.
[0030] Chloroquine antibody preparation:
[0031] Select healthy adult purebred BALA / C mice, take 50 μg of immune antigen prepared by coupling with protein and mix with the same amount of complete Freund's adjuvant, and use intraperitoneal injection for initial immunization, and then use the same dose of immune antigen plus the same amount every 3 weeks Incomplete Freund's adjuvant was used for the second and third immunizations by intr...
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The invention discloses preparation of an ELISA kit for detecting chloroquine and a detecting method of ELISA kit. The ELISA kit is sensitive, accurate and quick in detection, simple and convenient to operate and strong in specificity, and is suitable for batch sample detection. The kit comprises an elisa plate coated with chloroquine antigen, a chloroquine standard product, a chloroquine antibody working solution, a chloroquine enzyme-labeled second antibody working solution, a substrate solution A, a substrate solution B, a stopping solution, a concentrated sample diluting solution and a concentrated cleaning solution. The principle of the chloroquine detecting kit is solid-phase indirect competition ELISA, the extracted sample, enzyme-labeled second antibody working solution and antibody working solution are added into corresponding enzyme-labeled holes, after a period of incubation, the substrate solution A and the substrate solution B are added into a cleaning plate, the holes turn blue under the action of enzymes, then the stopping solution is added, and the color turns from blue into yellow. The shade of coloration is in inversely proportional relationship with the content of chloroquine in the standard product or the sample. The method can be directly used for detecting the content of chloroquine in feed and animal tissues.
Description
technical field [0001] The invention relates to the technical field of ELISA detection, in particular to an ELISA kit for detecting chloroquine. 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] At present, the methods for the determination of chloroquine mainly include high-performance liquid chromatography, reversed-phase high-performance liquid chromatography, and surface-enhanced Raman spectroscopy. The detection results are accurate and reliable, but the instruments and equipment are relatively expensive, and the sample pretreatment ...
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Inventor 洪霞邓榕榕杜霞
Owner JIANGSU WISE SCI & TECH DEV