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Enzyme linked immunosorbent assay kit for detecting sarafloxacin and detecting method thereof

A technology of enzyme-linked immunosorbent reagents and detection methods, which is applied in the field of enzyme-linked immunosorbent assay kits, can solve the problems of complex instrument setup, cumbersome process, unsuitability for on-site monitoring and screening of a large number of samples, and achieve sensitive detection and easy operation Effect

Inactive Publication Date: 2018-06-29
JIANGSU WISE SCI & TECH DEV
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  • Description
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Problems solved by technology

The chemical methods for detecting sarafloxacin residues mainly include thin layer chromatography (TLC), gas chromatography (GC), high performance liquid chromatography (HPLC), gas-mass spectrometry (GC / MS), liquid-mass spectrometry ( HPLC / MS), capillary electrophoresis (CE), etc., are not suitable for on-site monitoring and screening of a large number of samples due to complex instrument settings and cumbersome processes

Method used

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Embodiment Construction

[0021] Preparation of sarafloxacin protein conjugate:

[0022] The sarafloxacin hapten derivative with carboxyl group was obtained by the succinic anhydride method, and then 0.05 mmol and the carrier protein BSA were mixed in 0.05 mol / L carbonate buffer (CBS) with a pH of 9.6 at a binding ratio of 10:1. Then add 0.15mmol carbodiimide, stir at room temperature to react for 24 hours, and finally dialyze in 0.2 mol / L pH 7.6 PBS buffer for two days to remove unreacted hapten, and put the obtained protein conjugate solution in- Store at 20°C for later use.

[0023] Preparation of sarafloxacin antibody:

[0024] Choose healthy adult purebred BALA / C mice, take 50μg of immune antigen prepared by protein coupling and mix the same amount of complete Freund's adjuvant for the primary immunization, and then use the same dose of immune antigen every 3 weeks plus the same amount Incomplete Freund’s adjuvant was administered by intraperitoneal injection for the second and third immunizations. Aft...

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Abstract

The invention provides an enzyme linked immunosorbent assay kit for detecting sarafloxacin and a detecting method thereof. The enzyme linked immunosorbent assay kit for detecting sarafloxacin is sensitive to detect, accurate, quick, simple and convenient to operate, high in specificity, and suitable for detection of massive samples. The kit comprises an elisa plate coating sarafloxacin antigen, astandard sarafloxacin product, a first sarafloxacin antibody working solution, a second sarafloxacin enzyme-labelled antibody working solution, a substrate solution A, a substrate solution B, a stop solution, a concentrated diluting solution and a concentrated washing solution. The theory of the enzyme linked immunosorbent assay kit for detecting sarafloxacin is that solid phases indirect competeenzyme-linked immunosorbent assay; the extracted sample, the second enzyme-labelled antibody working solution and the antibody working solution are added to the corresponding micropores of the elisa plate and are incubated for a period of time; the substrate solution A and the substrate solution B are added to a washing plate; a developing agent is blue under the effect of enzyme; then the stop solution is added, and the color changes from blue to yellow; the developing level is in inverse proportion with the sarafloxacin content in a standard product or the sample. The method is directly applicable to the detection of residues of sarafloxacin in muscle tissues.

Description

Technical field [0001] The invention relates to the technical field of veterinary drug residue detection, in particular to an enzyme-linked immunoassay kit for detecting sarafloxacin in rice. Background technique [0002] Sarafloxacin is a fluoroquinolone antibiotic with the following characteristics: Due to its unique chemical structure, it has become a very important antibiotic in veterinary clinical medicine. Sarafloxacin has broad-spectrum antibacterial activity against gram-negative bacteria, gram-positive bacteria and mycoplasma pathogens. It also has a bactericidal effect on mycoplasma at a certain concentration, and it can also kill bacteria that are resistant to antibiotics. , Such as anti-p-lactic acid bacteria, amino glycosides and so on. Because of its unique effects, sarafloxacin has been widely used in the prevention and treatment of animal, fish and shrimp diseases since the 1980s. However, recent studies have found that all animals that have used sarafloxacin ha...

Claims

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Application Information

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IPC IPC(8): G01N33/543
CPCG01N33/543
Inventor 杜道林薛永来杜霞
Owner JIANGSU WISE SCI & TECH DEV
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