Immuomagnetic bead chromatographic test strip for rapidly detecting chloromycetin and preparation method thereof

A technology of immunomagnetic bead chromatography and detection test paper, which is applied in the direction of measuring devices, analytical materials, instruments, etc., can solve the problems of complicated preparation, tediousness, and damage to samples, and achieve the effects of high sensitivity, short reaction time, and cheap instruments and equipment

Inactive Publication Date: 2009-10-07
JIANGNAN UNIV
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  • Abstract
  • Description
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Problems solved by technology

At present, the detection methods of chloramphenicol drug residues adopted in my country include microbiological method, capillary zone electrophoresis, high performance liquid chromatography, gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, etc., but the above analysis methods have the following disadvantages: A series of physical or chemical operations will destroy the sample; the sample must be diluted, filtered and extracted in multiple steps, making the preparation complicated and cumbersome
There is no report on the development of rapid diagnostic test paper for the detection of chloramphenicol antibody by using nano-immunomagnetic beads

Method used

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  • Immuomagnetic bead chromatographic test strip for rapidly detecting chloromycetin and preparation method thereof
  • Immuomagnetic bead chromatographic test strip for rapidly detecting chloromycetin and preparation method thereof
  • Immuomagnetic bead chromatographic test strip for rapidly detecting chloromycetin and preparation method thereof

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preparation example Construction

[0027]2) Preparation and purification of polyclonal antibody: CAP-BSA was used as the immunogen, and New Zealand white rabbits were used as the immunized animals. For the first immunization, the immune source was mixed with an equal amount of Freund's complete adjuvant to make an emulsifier. Multi-point subcutaneous injection on the back of white rabbits, the immunization dose is 1 mg / rat. After 2-4 weeks, the same dose of immune source and the same amount of Freund's incomplete adjuvant are mixed to make an emulsifier, and the immunization is strengthened. During the immunization, the antibody effect is monitored. Valence and specificity, the last immunization without adjuvant. Seven days after the last immunization, the heart was bled, and purified polyclonal antibody to CAP was obtained by fractional precipitation with sulfuric acid. And the antibody was purified by ammonium sulfate precipitation method; rabbit anti-CAP-BSA IgG was obtained.

[0028] 3) Preparation of immu...

Embodiment 1

[0033] Detection of chloramphenicol CAP in aquatic products: Weigh 5g of the pulverized sample into a 50mL centrifuge tube, add 15mL of 80% acetonitrile aqueous solution to vortex, then ultrasonically extract, finally add 1g of sodium chloride to dissolve, and centrifuge at 3000rpm After 10 min, extract the supernatant organic phase, and then dry it with nitrogen. Finally, the residue after drying was dissolved with a buffer solution to obtain a sample solution to be tested. Finally, use the chloramphenicol immunomagnetic bead chromatography detection test strip to detect the sample solution. The insertion depth does not exceed the marked line. The sample will flow through the test strip through capillary chromatography. After 10 to 20 seconds, take out the test strip, After reacting at room temperature for 15 minutes, put the test strip into the Enviso TM The Magnetic Assay Reader of the System detects the magnetic field signal. According to the calibration curve measured ...

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Abstract

The invention discloses an immuomagnetic bead chromatographic test strip for rapidly detecting chloromycetin and a preparation method thereof, relating to the technical field of chloromycetin detection. The test strip consists of a lining board as well as a sample pad, a magnetic scale combination pad, a coated film and an absorbing pad which are sequentially engaged on the lining board; the magnetic scale combination pad is glass fiber for absorbing chloromycetin magnetic scale antibody; and the coated film is provided with an adelomorphic detection line T line which is printed by chloromycetin-coupled carrier protein solution, and an adelomorphic control line C line which is printed by goat anti-rabbit IgG solution. The sample pad of the test strip is inserted into the solution of a tested sample, then taken out after 10-20 seconds, and then reacts for 15 minutes at room temperature; then the test strip is arranged in a magnetic signal detector for detection, and the detector further outputs the biological responding signals which have been converted into magnetic field signals in the form of electrical signals; and after a specification curve is drawn, the chloromycetin content in the tested sample is calculated according to the specification curve. The invention relates to the method based on nano-magnetic immunological technique for detecting chloromycetin, and has the characteristics of high sensitivity, short reaction time, cheap instrument and equipment, convenience, easy usage and the like.

Description

technical field [0001] An immunomagnetic bead chromatography test strip for rapid detection of chloramphenicol and a preparation method thereof relate to the technical field of chloramphenicol detection. Background technique [0002] Chloramphenicol (Chloramphenicol, referred to as CAP) is the first fully chemical synthetic antibiotic, due to its low price, excellent antibacterial properties, and stable drug properties, it was once used as a feed additive and a therapeutic drug for bacterial diseases for a period of time. Wide range of use. Chloramphenicol is a broad-spectrum antibiotic. Repeated use of large doses will cause CAP to remain in milk and milk products. Chloramphenicol has a certain cumulative damage to the stomach, intestines, liver, etc., and can also cause allergic reactions, superinfection, teratogenicity, and easily cause human blood poisoning, leading to irreversible aplastic anemia. Therefore, all countries have set strict limits on the residual amount ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/552G01N33/532G01N33/558
Inventor 胥传来李灼坤刘丽强马伟
Owner JIANGNAN UNIV
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