Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Fluorescent probe for detecting ochratoxin A and preparation method thereof

A technology of ochratoxin and fluorescent probe, applied in the field of biochemistry, can solve the problems of complicated experimental steps, expensive instruments, and high costs

Active Publication Date: 2021-09-10
CHANGCHUN UNIV OF SCI & TECH
View PDF5 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above methods are limited by high cost, long period, and high technical requirements; or by complicated experimental steps, expensive instruments, time-consuming pretreatment, and inability to detect on-site.
It is worth noting that the core technology of immunoassay OTA is the preparation of OTA antibody, but the preparation of OTA monoclonal antibody is cumbersome and expensive

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Fluorescent probe for detecting ochratoxin A and preparation method thereof
  • Fluorescent probe for detecting ochratoxin A and preparation method thereof
  • Fluorescent probe for detecting ochratoxin A and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] The establishment of embodiment 1 OTA detection method

[0087] 1. Screening of specific affinity ligands (including non-specific panning and specific panning)

[0088] (1) The first round of non-specific elution

[0089] a. Use OTA with 0.1mol / L, pH 8.6 NaHCO 3 The solution was prepared as a solution of 80-100 μg / mL. Take 1 mL of this solution and coat it on a sterile polystyrene petri dish (60×15 mm), and carefully rotate repeatedly until the surface of the petri dish is completely wet. After completion, place the culture dish in a plastic box covered with wet gauze and incubate overnight at 4°C.

[0090] b. After overnight incubation, pour off the coating solution in the petri dish, and shake vigorously on the filter paper that has been irradiated by ultraviolet light to remove the residual coating solution. After removing the residual liquid, add 1 mL of blocking solution to the Petri dish, and block at 4°C for more than 1 hr.

[0091] c. After blocking, remove...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a fluorescent probe for detecting ochratoxin A and a preparation method thereof, and belongs to the technical field of biochemistry. The fluorescent probe is characterized in that OTA is taken as a target, and a phage surface random display 12-mer peptide library is used to screen out a recombinant phage specifically bound to the OTA. By extracting single-stranded DNA of the recombinant phage for sequencing and sequence comparison, an OTA specific affinity ligand sequence, Met-Pro-Met-Phe-Lys-His-Arg-Met-Phe-His-Thr-His is obtained. Afterwards the fluorescent probe specifically bound to OTA, FITC-Acp-Met-Pro-Met-Phe-Lys-His-Arg-Met-Phe-His-Thr-His is obtained by solid-phase polypeptide synthesis and fluorescence labeling. The construction method comprises the following steps: 1, screening a specific affinity ligand; 2, extracting recombinant phage DNA and determining a target sequence; 3, preparing a fluorescent probe; and 4, establishing a standard detection curve. The fluorescent probe of the present invention has the beneficial effects of 1, replacing an OTA monoclonal antibody; and 2, qualitatively identifying OTA and detecting content of the OTA in a sample.

Description

technical field [0001] The invention belongs to the field of biochemical technology. Background technique [0002] Ochratoxin is a class of toxic secondary metabolites produced by some genera of Penicillium and Aspergillus. Among them, ochratoxin A (Ochratoxin A, OTA) is the most toxic, widely distributed in various foods and crops including beer, wine, coffee beans, spices, vegetables, tea, fruit juice, bread, barley, wheat, corn, etc. It is also commonly found in feed. OTA is a mycotoxin second only to aflatoxin, which has a great impact on human and animal health. The main manifestations are carcinogenic, teratogenic, mutagenic, nephrotoxic, hepatotoxic, immunotoxic, etc. It is listed as a 2B carcinogen by the International Agency for Research on Cancer, which seriously threatens human health. The current methods for detecting OTA have certain defects. It is urgent to develop new detection reagents and establish new rapid, sensitive and specific detection methods to pr...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C07K1/13C40B30/04G01N33/533G01N33/53G01N33/58G01N33/68C09K11/06
CPCC07K7/08C40B30/04G01N33/533G01N33/5308G01N33/582G01N33/68C09K11/06C09K2211/1044Y02A50/30
Inventor 张淑华张馨予孙元宏罗杰张婷婷王思雅
Owner CHANGCHUN UNIV OF SCI & TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com