Method for quickly detecting enrofloxacin in meat chicken by nanometer biosensor based on immunomagnetic beads and fluorescent quantum dots

A technology of fluorescent quantum dots and biosensors, which is applied in the field of rapid detection of enrofloxacin in broilers by nano-biosensors, which can solve the problems of long detection time, complicated sample pretreatment process, and inapplicability for rapid detection.

Inactive Publication Date: 2019-01-29
ZHEJIANG UNIV
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  • Claims
  • Application Information

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Problems solved by technology

[0004] In the detection field of enrofloxacin, high performance liquid chromatography (HPLC) is currently the mainstream method for detecting enrofloxacin, which has high sensitivity and good accuracy, but requires complicated sample pretreatment process, and the detection time is long and The detection cost is high, so it is not suitable for on-site rapid detection
Enzyme-linked immunosorbent assay (ELISA) can greatly simplify the analysis process, with high specificity and relatively fast, but poor stability

Method used

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  • Method for quickly detecting enrofloxacin in meat chicken by nanometer biosensor based on immunomagnetic beads and fluorescent quantum dots
  • Method for quickly detecting enrofloxacin in meat chicken by nanometer biosensor based on immunomagnetic beads and fluorescent quantum dots
  • Method for quickly detecting enrofloxacin in meat chicken by nanometer biosensor based on immunomagnetic beads and fluorescent quantum dots

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Experimental program
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Effect test

Embodiment

[0079] 1) Preparation of IMBs modified by enrofloxacin monoclonal antibody

[0080] 1.1) Prepare 400 μL of a mixed solution of 10 mM EDC and 15 mM NHSS;

[0081] 1.2) Disperse carboxylated magnetic beads (purchased from Dayang Nano Technology Co., Ltd., USA) in 2-(N-morpholine)ethanesulfonic acid (MES) buffer at pH 6.0, and use MES buffer for magnetic separation and removal in sequence The treatment step of washing the supernatant was repeated 3 times, resuspended in 400 μL of the EDC and NHSS mixture described in step 1.1), and rotated and mixed at room temperature for 30 minutes to obtain an activated magnetic bead solution; the activated magnetic beads The solution was subjected to sequential magnetic separation and washing supernatant removal steps with pH 7.4 borate buffer and repeated 3 times, and then dispersed in 400 μL of borate buffer to obtain borate buffer containing activated magnetic beads. Dispersions;

[0082] 1.3) Mix 400 μL of borate buffer dispersion conta...

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Abstract

The invention discloses a method for quickly detecting enrofloxacin in meat chicken by nanometer biosensor based on immunomagnetic beads and fluorescent quantum dots. The surfaces of carboxylated magnetic beads are modified by enrofloxacin monoclonal antibodies, and sealed by bovine serum albumin, to prepare the solution of immunomagnetic beads sealed by the bovine serum albumin and modified by the enrofloxacin monoclonal antibodies; the enrofloxacin is coupled with the bovine serum albumin to prepare an enrofloxacin-bovine serum albumin compound; the acquired BSA-ENR compound is coupled withcarboxylated quantum dots to prepare a quantum dot-bovine serum albumin-enrofloxacin compound; a chicken tissue sample is processed to acquire chicken sample extracting solution; and the concentrationof the enrofloxacin in the extracting solution is quantitatively determined. The method provided by the invention is simple, quick, and high in sensitivity, does not need a complex sample pretreatment process, and has the capacity of quickly detecting the enrofloxacin on the scene and good development prospect.

Description

technical field [0001] The invention relates to a method for rapidly detecting enrofloxacin in broiler chickens with a nano-biosensor based on immunomagnetic beads (IMBs) and fluorescent quantum dots (QDs). Background technique [0002] Enrofloxacin (Enrofloxacin, ENR), also known as ethyl ciprofloxacin, is a fluoroquinolone antibiotic, which can achieve bactericidal effect by inhibiting the activity of DNA gyrase. It has the characteristics of strong penetrating power and is widely used in the treatment of diseases of aquatic products, livestock and poultry animals. However, the drug will cause potential toxic and side effects to the human body, and a large amount of use will induce bacterial resistance and indirectly endanger human health. Therefore, the European Union, Japan and my country have made clear regulations on the maximum residue limits (MRLs) of enrofloxacin in food of animal origin. my country stipulates that the maximum limit of the total amount of enroflox...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/94G01N33/543G01N33/531
CPCG01N33/531G01N33/54326G01N33/54346G01N33/9446
Inventor 李延斌沈亚芳何雅雯傅迎春
Owner ZHEJIANG UNIV
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