Detection gel column of lincomycin and detection method of lincomycin

A technology of lincomycin and a detection method, applied in the field of detection, can solve the problems of not being suitable for screening and detection of a large number of samples, not suitable for rapid detection, and lincomycin not suitable for rapid detection, etc.

Inactive Publication Date: 2017-11-24
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, instrumental methods such as chromatography-mass spectrometry have sufficient sensitivity for the analysis of most chemical pollutant residues, but the detection not only requires expensive analytical instruments, but also requires the operation level of technicians and a large number of pre-treatment reagents
Moreover, before the detection process, it is generally necessary to perform complex derivatization processing on the analytes. The pre-treatment process is complex, time-consuming, and inefficient, and is not suitable for screening and detection of a large number of samples.
High-performan

Method used

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  • Detection gel column of lincomycin and detection method of lincomycin
  • Detection gel column of lincomycin and detection method of lincomycin
  • Detection gel column of lincomycin and detection method of lincomycin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] Preparation of detection gel column for lincomycin

[0089] 1. Preparation of detection gel

[0090] (1) Gel carrier activation: take 1.0g of cyanogen bromide (CN-Br) activated agarose gel (Sepharose 4B) and swell it with 1mmol / L HCl, then fully wash it with 200mL HCl solution, the whole process takes 15min completed within. The swollen gel was re-used with 0.1mol / L NaHCO 3 The solution is well equilibrated. Generally, 1.0g of Sepharose 4B activated by cyanogen bromide can swell about 3.5mL of gel.

[0091] (2) Coupling: Dissolve 0.3mL of the first polyclonal secondary antibody (goat anti-mouse antibody, secondary antibody, 2.3g / L) in 5mL of gel coupling solution, mix with the above-mentioned activated gel carrier, and stir for 20h .

[0092] (3) Washing: 50 mL of gel-coupling solution was used to wash to remove unbound primary polyclonal secondary antibody (goat anti-mouse antibody).

[0093] (4) Blocking: 20 mL of gel blocking solution was added to the gel solut...

Embodiment 2

[0111] Preparation of HRP-labeled lincomycin enzyme-labeled antigen (LIN-HRP)

[0112] Use 0.3mL ultrapure water (ddw) and 0.2mL N,N-dimethylformamide (DMF) mixture, DMF:H in the mixture 2 O=2:3, 2.7mg NaIO 4Dissolved in the mixture to obtain NaIO 4 solution. NaIO 4 The solution was added dropwise into 5.8 mg LIN dissolved in 0.2 mL of N,N-dimethylformamide (DMF), stirred at room temperature for 20 min, and this was liquid A. Weigh 3.35mg of HRP, use 0.13mol / L of NaHCO 3 The solution (2.76mL) was made into a 10% HRP activation solution, which was solution B. Add liquid B to liquid A dropwise, and stir for 2 h in the dark to obtain an intermediate product. Then add 0.1 mL of NaBH to the intermediate 4 (2mg / mL) reduction 2h. The synthesized lincomycin enzyme-labeled antigen was stirred and dialyzed with PBS at 4°C for 3 days, the medium was changed 4 times a day, and the purified LIN-HRP was obtained by centrifugation to remove the precipitate. The reaction equation is a...

Embodiment 3

[0115] The detection gel column (visualized gel ELISA detection gel column) of lincomycin prepared in Example 1 is used to detect the sample to be tested

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Abstract

The invention relates to a detection gel column of lincomycin and a detection method of lincomycin. The detection gel column of lincomycin comprises an open tubular column as well as a detection layer and a quality control layer which are filled in the open tubular column. During detection, a sample to be measured and a lincomycin enzyme-labelled antigen labelled with a chemiluminescent label are added into the detection gel column of lincomycin and flow through the detection layer and the quality control layer. The sample to be measured and the lincomycin enzyme-labelled antigen labelled with the chemiluminescent label together compete for a lincomycin monoclonal antibody in the detection layer, and color of the detection layer and color of the quality control layer are used for judging whether the sample to be measured contains lincomycin. The detection gel column of lincomycin has carrying convenience, and can be used for rapidly and sensitively detecting whether the sample to be measured contains lincomycin, and the method is suitable for rapid on-site detection of a lot of samples.

Description

technical field [0001] The invention relates to the technical field of detection, in particular to a lincomycin detection gel column and a lincomycin detection method. Background technique [0002] Lincomycin (Lincomycin, LIN), also known as lincomycin, is a lincomycin narrow-spectrum antibiotic obtained from the culture medium of Streptomyces. In veterinary clinics, lincomycin is mainly used to treat various infections caused by Gram-positive bacteria, especially penicillin-resistant Gram-positive bacteria, chronic respiratory diseases of poultry caused by Mycoplasma, swine panting, anaerobic Bacterial infections such as necrotic enteritis in chickens, etc., are also used to treat Treponema dysentery, toxoplasmosis and actinomycosis in dogs and cats. Lincomycin has outstanding curative effect on cow mastitis caused by Gram-positive bacteria and anaerobic bacteria, and can be used for systemic or local treatment. In the production of bee industry, it is mainly used for the...

Claims

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Application Information

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IPC IPC(8): G01N33/94G01N33/577
CPCG01N33/577G01N33/9446
Inventor 蒋文晓陈献雄柯跃斌翟鹏吕子全
Owner SHENZHEN UNIV
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