A method for the detection of multiple residues of antibiotics with an aptamer-based magnetic nanofluorescence sensor

A fluorescent sensor and aptamer technology, which is applied in the fields of inspection and quarantine and food safety, can solve the problems of cumbersome pre-treatment and inability to realize rapid on-site detection, and achieve the effects of shortening detection time, satisfying rapid detection and simple operation.

Active Publication Date: 2016-03-02
XINJIANG ACADEMY OF AGRI & RECLAMATION SCI
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AI Technical Summary

Problems solved by technology

Aiming at the shortcomings of traditional large-scale instruments that cannot realize on-site rapid detection, cumbersome pretreatment, and the need to prepare antibodies for immunoassay methods, the present invention establishes a fluorescence detection method based on magnetic bead separation-aptamer recognition, which can be used for oxytetracycline and oxytetracycline in food. Rapid and highly sensitive detection of multiple residues of kanamycin

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  • A method for the detection of multiple residues of antibiotics with an aptamer-based magnetic nanofluorescence sensor
  • A method for the detection of multiple residues of antibiotics with an aptamer-based magnetic nanofluorescence sensor
  • A method for the detection of multiple residues of antibiotics with an aptamer-based magnetic nanofluorescence sensor

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Embodiment 1

[0030] Example 1: Establishment of standard curve

[0031] The method in the content of the invention is used to determine the standard substance of oxytetracycline and kanamycin to establish a standard curve. The series of standard concentrations of oxytetracycline and kanamycin are respectively configured as 0, 0.5, 1, 2, 5, 10, 20, 50, 100, 200 ng / mL, with different standard concentrations as the abscissa and relative fluorescence values As the ordinate, draw a standard curve and establish a regression equation. The linear ranges of oxytetracycline and kanamycin are 1-100ng / mL and 2-100ng / mL, respectively.

Embodiment 2

[0032] Example 2 Milk sample detection

[0033] Purchase 5 portions of milk, measure 5.0 mL of milk sample, add 5 mL of 1% trichloroacetic acid solution, vortex to mix, and centrifuge at 10000r / min for 10 minutes. The upper fat layer was discarded, 200 μL of the clear liquid was distilled to 1 mL with 10 mmol / L PBS buffer, and the detection was carried out according to the detection method in the content of the invention, and the detection result was quantitatively analyzed according to the standard curve. Oxytetracycline was detected in 1 of the 5 samples, and the content was 76.2ng / ml, which was detected by the GB / T22990-2008 method. The detection value of this sample was 71.8ng / ml. All kanamycin was not detected. A sample was selected for the kanamycin addition and recovery experiment. The added concentration was 100ng / ml. The measured value of the method of the present invention was 80.0ng / ml, and the GB / T22969-2008 measured value was 85.3ng. / ml.

Embodiment 3

[0034] Example 3 Detection of animal tissue samples

[0035] Five samples of pork were purchased from the market, each weighing 2.0 g into a 50 mL centrifuge tube, adding 10 mL of 1% trichloroacetic acid solution, vortexing for 1 min, ultrasonic extraction for 5 min, and centrifugation at 10,000 r / min for 10 min. Take 250uL of the supernatant and dilute with 10mmol / LPBS buffer to 1mL for analysis, and calculate the content of the analyte in the sample according to the standard curve. Oxytetracycline and kanamycin were not detected in the 5 samples. One sample was selected for the addition and recovery experiment of oxytetracycline and kanamycin. The addition amount was 100ng / g. The detection value of oxytetracycline in the present invention 85.4ng / g, the detection value of kanamycin is 87.9ng / g, and the detection value of oxytetracycline according to GB / T20764-2006 is 90.2ng / g. There is no corresponding national standard for kanamycin without comparison.

[0036] The recovery rate...

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Abstract

The invention relates to a method for detecting multi-residues of antibiotics by a magnetic nano fluorescence sensor based on an aptamer, belonging to the field of food safety analysis. According to the method, terramycin and kanamycin aptamers are simultaneously modified on the surface of a nano magnetic micro-sphere; complementary chains of fluorescence radical marks are respectively combined by base complementation; when an object to be detected exists, the aptamer is combined with the object to be detected to cause complementary chain dissociation to change fluorescence intensity, so that the aim of quantitatively detecting terramycin and kanamycin is realized. The method utilizes a magnetic nano material to rapidly separate the target to be detected in a sample so that the detection time is shortened, the detection cost is reduced, the detection sensitivity is improved, the operation is simple and the aim of analyzing the multi-residues is realized.

Description

Technical field [0001] The invention relates to the detection of oxytetracycline and kanamycin using aptamer technology, and belongs to the technical field of food safety, inspection and quarantine. Background technique [0002] In recent years, food safety incidents have occurred frequently, seriously affecting consumer confidence. Among them, antibiotic residues have become the focus of attention. In the breeding process, tetracycline antibiotics and kanamycin are often used as antibacterial growth-promoting additives and therapeutic drugs. Due to improper use and non-compliance with the withdrawal period regulations, the residues in animal products may cause allergic reactions, disrupt the balance of gastrointestinal flora, and cause intestinal dysfunction and enhancement after being absorbed by the human body through the food chain. Bacterial drug resistance brings serious harm to human health. [0003] Currently, antibiotic detection methods mainly include high performance ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
Inventor 卢春霞唐宗贵罗小玲刘长彬王远贺玉凤王国红罗志根刘丽刘成江
Owner XINJIANG ACADEMY OF AGRI & RECLAMATION SCI
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