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Fluorescent sensor for detecting lipopolysaccharide, preparation method and application thereof

A fluorescent sensor and lipopolysaccharide technology, applied in fluorescence/phosphorescence, instruments, measuring devices, etc., can solve the problems of false positives in enzyme-linked immunoassay, long time-consuming rapid silver staining, and many false positive reactions, etc., and achieve the measured data Accurate and reliable, broaden the application, enhance the effect of anti-interference

Active Publication Date: 2019-08-02
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these methods have limitations to varying degrees. Among them, the limulus reagent method has many false positive reactions, the enzyme-linked immunosorbent assay is prone to false positive reactions, and the rapid silver staining method has defects such as time-consuming and cumbersome operations.

Method used

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  • Fluorescent sensor for detecting lipopolysaccharide, preparation method and application thereof
  • Fluorescent sensor for detecting lipopolysaccharide, preparation method and application thereof
  • Fluorescent sensor for detecting lipopolysaccharide, preparation method and application thereof

Examples

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Effect test

Embodiment 1

[0041] Embodiment 1: In this embodiment, see Figure 1 to Figure 7 , a fluorescent sensor for detecting lipopolysaccharide, mainly composed of a kit, a solid-liquid separation device, a fluorescence spectroscopic analysis device, an analysis system and a control system, the control system controls the supply of reagents in the kit and the input of detection data and output; the lipopolysaccharide aptamer (LPSA chain) is modified onto the magnetic beads (MB) by combining biotin and streptavidin, and a solid-liquid separation device is used to obtain the lipopolysaccharide aptamer covering the surface of the magnetic beads The magnetic beads (MB-LPSA), and then add the recognition strand DNA (L' strand) that can be complementary to the lipopolysaccharide aptamer base pairing, and obtain the magnetic beads (MB-LPSA / L') solution, forming a recognition system as the first reaction reagent, the modified lipopolysaccharide aptamer chain (LPSA) and the recognition strand DNA ( L') w...

Embodiment 2

[0063] This embodiment is basically the same as Embodiment 1, especially in that:

[0064] In this example, see Figure 5 and Figure 6 , for the detection of lipopolysaccharide at different concentrations.

[0065] Double-strand modified magnetic beads (MB-LPSA / L') were mixed with different concentrations of lipopolysaccharide (10 -4 ng / mL~10 7 ng / mL) reaction, after specific recognition binding and DNA walker reaction, it was detected by fluorescence spectrometry.

[0066] Such as Figure 5 As shown, the fluorescence intensity value of the reaction product solution at a wavelength of 520nm increases with the increase of the concentration of lipopolysaccharide. Figure 5It is the spectrogram in the wavelength range of 510nm to 600nm under the condition of the presence of various concentrations of lipopolysaccharide. Figure 6 It is a linear relationship graph between the 520nm fluorescence intensity value and the lipopolysaccharide concentration, at 10 -4 ng / mL~10 7 W...

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Abstract

The invention discloses a fluorescent sensor for detecting lipopolysaccharide, a preparation method and application thereof. The fluorescent sensor consists of a first reagent and a second reagent, wherein the first reagent is magnetic bead aqueous solution covered with lipopolysaccharide aptamer (LPSA) chains and recognition chain DNA which is in complementary base pairing with the lipopolysaccharide aptamer; the second reagent is a DNA Walker system formed by magnetic beads covered with long chains and short chains. A walker product is analyzed by a fluorescence spectroscopic technology, sothat analysis and detection of the lipopolysaccharide are realized. The fluorescent sensor of the invention has the advantages of simple method, stability, strong specificity, high sensitivity, capacity for linearly detecting the lipopolysaccharide within the range of 10<-4>ng / mL-10<7>ng / mL, and capacity for effectively distinguishing the lipopolysaccharide from other contrast substances.

Description

technical field [0001] The invention relates to a detection method and application of lipopolysaccharide, in particular to a fluorescent sensor for detection of lipopolysaccharide, its preparation method and application, which are applied in the technical fields of food quality control and health care management analysis. Background technique [0002] Lipopolysaccharide (LPS), a component of the cell wall of Gram-negative bacteria, is toxic to the host. Only when the bacteria die and dissolve or the bacterial cells are destroyed by artificial methods, the lipopolysaccharide is released, so it is also called endotoxin. The main toxic component of lipopolysaccharide is lipid A, which can cause fever, microcirculation disturbance, endotoxin shock and disseminated intravascular coagulation. It is heat-resistant and stable, and has weak antigenicity, but the human body is extremely sensitive to lipopolysaccharide. Studies have reported that a very small amount (1-5 ng / kg body w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N21/64
CPCG01N33/54326G01N21/6428G01N2021/6417
Inventor 张娟岳禧泉薛添香
Owner SHANGHAI UNIV