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Biosensor based on earthworm coelomic fluid, application of biosensor in detection of tetracycline antibiotics and detection method

A tetracycline and antibiotic technology, which is applied in the field of biosensors and achieves the effects of simple extraction method, good optical performance and simple detection process

Inactive Publication Date: 2021-08-31
四川中科微纳科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, various fluorescent probes have been synthesized for the detection of tetracycline antibiotics, but there is no research report on the use of fluorescent substances extracted from earthworms for the detection of tetracycline antibiotics

Method used

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  • Biosensor based on earthworm coelomic fluid, application of biosensor in detection of tetracycline antibiotics and detection method
  • Biosensor based on earthworm coelomic fluid, application of biosensor in detection of tetracycline antibiotics and detection method
  • Biosensor based on earthworm coelomic fluid, application of biosensor in detection of tetracycline antibiotics and detection method

Examples

Experimental program
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Effect test

Embodiment 1

[0037] A method for preparing a biosensor based on earthworm coelom fluid, comprising the following steps:

[0038] (1) Mix 10% ethanol, 0.01MPBS buffer solution and 2.5g / L ethylenediaminetetraacetic acid (EDTA) to prepare stimulation solution.

[0039] (2) Weigh, wash and dry the adult Eisenia chinensis, and put them into a Petri dish with a stimulating solution equal in volume to that of the earthworm.

[0040] (3) Collect the discharged body cavity fluid into a centrifuge tube with a pipette gun, and freeze at -80°C for later use.

Embodiment 2

[0042] The earthworm coelom fluid biosensor extracted and prepared in Example 1 was observed using laser confocal microscopy and flow cytometry ( Figure 1-3 ). The results showed that the coelom fluid of earthworms contains two kinds of cells, among which only granulosa cells have autofluorescence, and the fluorescence intensity is proportional to the size of the vesicles. Under different excitations, they can emit purple, green, yellow and red fluorescence, indicating that it has the ability to excite. dependency.

Embodiment 3

[0044] The biosensor of the earthworm coelom fluid prepared in Example 1 is used to detect tetracycline antibiotics, comprising the following steps:

[0045] (1) Prepare a 250 μmol / L standard solution of tetracycline antibiotics first, dilute the tetracycline antibiotics with PBS buffer (pH=7.2-7.4) and double distilled water to the following concentrations: 0.5, 0.75, 1, 2.5, 5, 7.5, 10, 25, 50, 75 μmol / L.

[0046] (2) Take out the earthworm coelom fluid frozen at -80°C and centrifuge for 10 min at 10,000 r / min, then collect the upper layer solution as the earthworm coelom fluid detection solution.

[0047] (3) Mix and shake 1mL of the above-mentioned tetracycline antibiotic solutions with different concentrations and 1mL of earthworm body cavity fluid detection solution, and start to scan the fluorescence spectrum at different excitation wavelengths after 1min, and measure the fluorescence spectrum in the range of 450-650nm to obtain the Optimal emission wavelength correspo...

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Abstract

The invention discloses a biosensor based on earthworm coelomic fluid, application of the biosensor in detection of tetracycline antibiotics and a detection method. The biosensor is derived from a coelomic fluid of eisenia foetida, wherein the coelomic fluid emits green fluorescence under excitation of 370 nm, and a fluorescence emission peak is located at 520 nm and can be quenched under the action of tetracycline antibiotics. The invention also discloses an application of the biosensor in selective detection of tetracycline antibiotics. In the detection process, along with the increase of the concentration of the added tetracycline antibiotics, the fluorescence intensity of an emission peak at 520 nm is reduced, so that the content of the tetracycline antibiotics in the sample can be detected according to the quenching degree of fluorescence. Compared with the traditional method, the method for detecting the tetracycline antibiotics provided by the invention has the advantages of high selectivity, high sensitivity, biological non-toxicity and compatibility.

Description

technical field [0001] The invention belongs to the technical field of biosensors, and in particular relates to a biosensor based on body cavity fluid of earthworms, a preparation method thereof, and a method for detecting tetracycline antibiotics by the biosensor. Background technique [0002] Tetracycline antibiotics are a class of broad-spectrum antibiotics. They usually have good antibacterial and sterilizing effects on most bacteria, so they are widely used to treat various diseases. They can also be used as veterinary drugs to promote animal growth and prevent disease etc. However, in recent years, the illegal abuse of tetracycline antibiotics in the livestock and poultry breeding process has been very serious, resulting in them being flooded in animal-derived foods. In addition, in the process of livestock and poultry farming, some tetracycline antibiotics cannot be absorbed by animals, and are excreted in the form of mothers, accumulate in soil and freshwater enviro...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/64G01N21/6428G01N2021/6417G01N2021/6432
Inventor 霍峰蒋志刘冠辉王曦妮王显祥
Owner 四川中科微纳科技有限公司
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