Method for detecting ochratoxin A by magnetic separation of aptamer-functionalized magnetic nano material and marking of up-conversion fluorescent nano material

A technology of fluorescent nanomaterials and ochratoxin, which is used in material inspection products, material excitation analysis, fluorescence/phosphorescence, etc., can solve the problems of complicated separation steps, affecting detection sensitivity and accuracy, etc., to shorten the detection period and improve the stability. Performance and accuracy, simple operation

Active Publication Date: 2011-04-20
JIANGNAN UNIV
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Problems solved by technology

At the same time, in the detection of actual biological samples, due to the small number of samples to be tested and the complicated separation steps in labeling and

Method used

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  • Method for detecting ochratoxin A by magnetic separation of aptamer-functionalized magnetic nano material and marking of up-conversion fluorescent nano material
  • Method for detecting ochratoxin A by magnetic separation of aptamer-functionalized magnetic nano material and marking of up-conversion fluorescent nano material
  • Method for detecting ochratoxin A by magnetic separation of aptamer-functionalized magnetic nano material and marking of up-conversion fluorescent nano material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Establishment and detection of ochratoxin A detection standard curve in actual corn samples

[0027] Sample pretreatment: crush and sieve wheat at high speed, weigh 20g into a 100mL flask, add 5g NaCl, 80% ethanol aqueous solution, mix thoroughly, place in a homogenizer for high-speed stirring and extraction for 2min, stand still for a while, filter, and take 10mL filtrate Put it in a 50mL flask, stir and extract again in a homogenizer at high speed for 2min, after resting, filter with ultra-fine glass fiber filter paper until the filtrate is clear, collect the filtrate for later use.

[0028] Purchased 16 kinds of different types of wheat from six local farmer's markets, and used the method of the present invention and the national standard method to measure the content of ochratoxin A in it respectively. both Significant differences. It shows that the inventive method is fast and reliable, has high sensitivity and good stability, and is suitable for the ...

Embodiment 2

[0032] Example 2: The detection of ochratoxin A in the actual corn sample and the recovery rate of the standard addition The experimental sample pretreatment is the same as that in Example 1.

[0033] Select three groups from the 16 groups of ochratoxin A concentration data that embodiment 1 obtains as background value, then select 0.5 * 10 -12 g / mL, 1.0×10 -12 g / mL, 5.0×10 -12 OTA standard substances with three different concentrations of g / mL were added to the test substance respectively, and the content of OTA therein was detected again by the method of the present invention to obtain the detection value. Recovery %=(detection value-background value) / addition amount×100%. From the results in Table 2, the recovery rate is 90.70%-117.98%, indicating that the present invention is stable, sensitive and accurate, and is suitable for the detection of ochratoxin A in actual corn samples.

[0034] Table 2: Detection and recovery of ochratoxin A in real corn samples

[0035]

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Abstract

The invention discloses a method for detecting ochratoxin A by the magnetic separation of an aptamer-functionalized magnetic nano material and the marking of an up-conversion fluorescent nano material, which is used for detecting the ochratoxin A content of wheat, grains, feeds and products thereof, and the like. In the method, the ochratoxin-specific aptamer-functionalized magnetic nano material is utilized to perform magnetic separation and enrichment reaction on a sample to rapidly condense bulk samples, effectively increase the condensation of the samples by thousands of times, greatly shorten a detection period and improve the detection sensitivity; and simultaneously, the novel ochratoxin A analysis method for realizing rapid sample treatment, analysis and detection is constructed by combining the characteristics of high sensitivity of laser induced fluorescence of the up-conversion fluorescent nano material and capability of effectively avoiding the interference of biological background fluorescence of the samples.

Description

technical field [0001] The invention relates to the detection of ochratoxin A by using magnetic nanomaterials and up-conversion fluorescent nanomaterials combined with nucleic acid aptamer recognition, and is characterized in that the magnetic nanomaterials functionalized by ochratoxin A aptamers are used to detect ochratoxin in bulk samples The separation and enrichment of A shortens the detection period and improves the detection sensitivity. At the same time, the up-conversion fluorescent nanomaterial is used as a marker to further improve the detection sensitivity by detecting the laser-induced up-conversion fluorescence signal. The method belongs to the field of nanomaterials and molecular biotechnology. Background technique [0002] Ochratoxin A (ochratoxinA, OTA) is a space metabolite produced by certain species of Aspergillus and Penicillium. Mutagenesis and other toxicity have great potential harm to animal and human health. OTA is widely distributed in nature, and...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N33/68
Inventor 王周平吴世嘉段诺
Owner JIANGNAN UNIV
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