Eureka AIR delivers breakthrough ideas for toughest innovation challenges, trusted by R&D personnel around the world.

Detection kit of ochratoxin and method thereof for detecting ochratoxin

A technology of ochratoxin and detection kit, which is applied in the direction of measuring devices, instruments, fluorescence/phosphorescence, etc., can solve the problems of low detection sensitivity and inconvenient operation of ochratoxin, achieve high sensitivity and specificity, rapid detection and easy operation simple effect

Active Publication Date: 2019-07-19
QINGDAO AGRI UNIV
View PDF6 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In view of the problems existing in the prior art, the object of the present invention is to provide a kit for detecting ochratoxin and a method for detecting ochratoxin, so as to alleviate the low detection sensitivity and inconvenient operation of ochratoxin in the prior art. technical issues

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
  • Detection kit of ochratoxin and method thereof for detecting ochratoxin
  • Detection kit of ochratoxin and method thereof for detecting ochratoxin
  • Detection kit of ochratoxin and method thereof for detecting ochratoxin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Embodiment 1 is used for the test kit that detects ochratoxin

[0056] This embodiment provides a kit for detecting ochratoxin, which includes the following substances:

[0057] The nucleic acid aptamer of ochratoxin has the sequence shown in SEQ ID NO.1:

[0058] 5'-GATCGGGTGTGGGTGGCGTAAAGGGAGCATCGGACA-3' (SEQ ID NO.1);

[0059] The cDNA that is not completely complementary to the ochratoxin nucleic acid aptamer has the sequence shown in SEQ ID NO.2:

[0060] 5'-TGTCCGTAGCTCCCAATTCACCTCCGTCACCCGATC-3' (SEQ ID NO.2);

[0061] The hairpin sequence H1 has the sequence shown in SEQ ID NO.3:

[0062] 5'-AGGGCGGGGGTGTGGGGGCGTCTAACGGAGGTGAATTGGGTTT-3' (SEQ ID NO. 3);

[0063] The hairpin sequence H2 has the sequence shown in SEQ ID NO.4:

[0064] 5'-TGGGTAGACGCCCCCACACCAAGAAGAAGAATGCCTCCACTTAACCCTA-3' (SEQ ID NO.4)

[0065] Hemoglobin;

[0066] Tetramethylbenzidine (TMB).

[0067] Among them, the ochratoxin nucleic acid aptamer binds incompletely to the cDNA, and H1 a...

Embodiment 2

[0068] Embodiment 2 detects the method for ochratoxin

[0069] This embodiment provides the method for using the kit provided in Example 1, comprising the following steps:

[0070] (1) After mixing the ochratoxin nucleic acid aptamer and the cDNA that is not completely complementary to the nucleic acid aptamer, put it in a water bath at 95°C for 5 minutes, and then cool it naturally at room temperature, so that the two can form a double strand through nucleic acid complementary pairing;

[0071] (2) Add 2 μL of PBS to the system to dilute the ochratoxin test sample, and react at room temperature for 30 minutes, so that the ochratoxin binds to the ochratoxin nucleic acid aptamer in the double strand and opens the double strand structure; The cDNA that is not completely complementary to the ligand dissociates into a single strand;

[0072] (3) Add the mixed solution of hairpin sequences H1 and H2 to the reaction system in 2), react at room temperature for 1 hour, and carry out ...

Embodiment 3

[0075] The detection of the target ochratoxin of embodiment 3 different concentrations

[0076] In this embodiment, the target ochratoxins of different concentrations were detected, specifically including the following:

[0077] Prepare standard solutions of ochratoxin with final concentrations of 10 μg / mL, 100 μg / mL, 500 μg / mL and 1 mg / mL and store at room temperature.

[0078] Different concentrations of ochratoxins were detected according to the method in Example 2, reacted fully, detected the absorbance intensity at 450nm and observed the color change with naked eyes.

[0079] from figure 2 It can be seen from the detection results that when ochratoxin is present, the color changes from colorless to blue (not shown in the figure), and the absorbance value increases significantly; and as the concentration of ochratoxin increases, the absorbance value increases. Among them, the Hemin blank group is only hemin plus magnetic beads, without adding the DNA product of the prev...

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 detection kit of ochratoxin and a method thereof for detecting the ochratoxin, and belongs to the technical field of detection of harmful substances. The detection kit of ochratoxin disclosed by the invention comprises a nucleic acid aptamer of the ochratoxin, a cDNA which is not completely complementary to the nucleic acid aptamer, and two hairpin sequences H1 and H2, wherein the sequence of the nucleic acid aptamer of the ochratoxin is as shown in a sequence table SEQ ID NO. 1; the two hairpin sequences H1 and H2 can be self-assembled into a long chain; and the cDNAwhich is not completely complementary to the nucleic acid aptamer of the ochratoxin can trigger the self-assembly of the hairpin sequences H1 and H2. The kit provided by the invention has very high sensitivity and specificity, simple operation and rapid detection, the result is visible to naked eyes, the kit is suitable for promotion in all laboratories, and real-time onsite analysis of the ochratoxin can be realized without professional training.

Description

technical field [0001] The invention belongs to the technical field of detection of harmful substances, and in particular relates to a detection kit for ochratoxin and a method for detecting ochratoxin. Background technique [0002] Ochratoxins are natural contaminants belonging to the mycotoxin group, produced by various fungal types Aspergillus and Penicillium. Ochratoxin A (OTA), a mycotoxin, was discovered in South African corn flour in the 1960s. Years of studies have shown that OTA has nephrotoxicity, hepatotoxicity, embryotoxicity, teratogenicity, neurotoxicity, immunotoxicity, genotoxicity and carcinogenicity. Of the 6 known ochratoxin types, ochratoxin A exhibits the highest toxicity. The International Agency for Research on Cancer (IARC) has classified OTA as a probable human carcinogen (category 2B) based on substantial evidence of carcinogenicity found in several animal studies. Because OTA exists widely in nature and is very stable and not easy to metabolize,...

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
IPC IPC(8): G01N21/64
CPCG01N21/6486
Inventor 吴薇杨庆利于春娣
Owner QINGDAO AGRI UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
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