Probe group and kit for detecting aflatoxin B1 and application of probe group and kit

An aflatoxin and probe set technology, applied in the biological field, can solve the problems of complicated operation, time-consuming and labor-intensive, high cost of aflatoxin detection, etc., and achieve the effect of improving detection sensitivity and low cost

Active Publication Date: 2021-03-30
JIANGSU INST OF NUCLEAR MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Therefore, the technical problem to be solved by the present invention is to overcome the defects of high cost, cumbersome operation, time-consuming and labor-consuming and difficult to popularize the detection of aflatoxins in traditional Chinese medicines in the prior art, thereby providing a simple, sensitive and low-cost label-free Probe set, kit, sensor system and detection method for detecting aflatoxin B1

Method used

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  • Probe group and kit for detecting aflatoxin B1 and application of probe group and kit
  • Probe group and kit for detecting aflatoxin B1 and application of probe group and kit
  • Probe group and kit for detecting aflatoxin B1 and application of probe group and kit

Examples

Experimental program
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Effect test

experiment example 5

[0083]When testing the actual sample in Experimental Example 5, the sample to be tested is 1 μL of 100 ng / mL (or 1 μL of 10 ng / mL, or 1 μL of 1 ng / mL) AFB1 solution. Here, for the preparation of three concentrations of AFB1 solutions, the solvent used is the actual sample extract instead of benzene:methanol (98:2, volume ratio) solvent. Taking the 100ng / mL AFB1 solution as an example, the preparation process of the sample to be tested is as follows: pipette 0.5 μL of 2.0 μg / mL AFB1 standard, and add 9.5 μL of the actual sample extract. Available after dilution, 10 μL of 100ng / mL AFB1 solution. Other concentrations of AFB1 were diluted step by step in the same way to AFB1 solutions with concentrations of 10ng / mL and 1ng / mL.

[0084] The room temperature range in the following examples is 10-30°C.

Embodiment 1

[0085] Embodiment 1 probe set

[0086] 1.1 Probe set, including:

[0087] The aptamer probe H1-S1 is formed by hybridizing the hairpin probe H1 and the aptamer sequence S1 for binding the target; the hairpin probe H1 includes a loop region and a stem region, and the loop region is Part of the G-quadruplex sequence, and the remaining part of the G-quadruplex sequence is located in the stem region; the 5' end of the hairpin probe H1 has a protruding single-stranded DNA sequence 1, and the hairpin probe 1 has a protruding single-stranded DNA sequence 1. The 3' end has a protruding single-stranded DNA sequence 2; the DNA sequence 1 and DNA sequence 2 respectively hybridize complementary to the sequences at both ends of the aptamer sequence S1 and form blunt ends;

[0088] The hairpin probe H2 includes a loop region and a stem region, the loop region is a part of the G-quadruplex sequence, and the remaining part of the G-quadruplex sequence is located in the stem region; the hairp...

Embodiment 2

[0097] Embodiment 2 detects the kit of aflatoxin B1

[0098] This embodiment provides a test kit for detecting aflatoxin B1, comprising:

[0099] Aptamer probes H1-S1 and hairpin probe H2 prepared in Example 1.

[0100] Further, the kit also includes independently packaged exonuclease I.

[0101] Further, it also includes individually packaged hemin, luminol and / or H 2 o 2 .

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Abstract

The invention relates to the technical field of biology, in particular to a probe set, a kit and a sensor system for detecting aflatoxin B1 and application of the probe set, the kit and the sensor system. The invention successfully constructs a probe group for high-sensitivity detection of AFB1 based on an enzyme-assisted target cyclic amplification strategy. The designed aptamer probe H1S1 and hairpin probe H2 are high in AFB1 detection sensitivity, wide in linear range and good in specificity; and the probe group takes the Gquadruplex / hemin DNA enzyme as a signal molecule, does not need anychemical modification and fluorescence labeling, and is low in preparation cost. When the probe group is used for detecting the target AFB1, the whole amplification reaction is carried out in an isothermal uniform solution, a complex separation procedure and time-consuming thermal cycle are not needed, and the operation is simple.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a probe group, a kit, a sensor system and an application thereof for detecting aflatoxin B1. Background technique [0002] Aflatoxin (AFT) is a kind of toxin with difuran ring and coumarin structure produced by fungi such as Aspergillus flavus and Aspergillus parasiticus, among which aflatoxin B1 (AFB1) is the most toxic and the most carcinogenic currently known. It is a strong natural toxin and was classified as a class 1 carcinogen by the International Agency for Research on Cancer (IARC) in 1993. In the process of production, processing, storage, transportation, etc., Chinese herbal medicines are prone to mildew and aflatoxin contamination due to changes in temperature or humidity. The 2020 edition of the "Chinese Pharmacopoeia" contains the limited detection of aflatoxin residues in 24 kinds of Chinese medicinal materials, and stipulates that the content of AFB1 shall not exceed...

Claims

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Application Information

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IPC IPC(8): C12N15/115G01N33/53C12Q1/682G01N21/76
CPCC12N15/115G01N33/53C12Q1/682G01N21/76G01N2333/38C12N2310/16
Inventor 吴昊邹霈吴军王洪勇刘娅灵韩国庆李文超
Owner JIANGSU INST OF NUCLEAR MEDICINE
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