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A technology of gold seeds and magnetic beads, which is applied in biochemical equipment and methods, measuring devices, microbiological determination/inspection, etc., can solve the problems of false positive reactions and achieve false positive reaction avoidance, low cost and high sensitivity operation Effect
Inactive Publication Date: 2019-12-20
FUZHOU UNIV
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However, most SERS sensors are used to detect OTA in the "off" mode, and there may be false positive reactions in the detection process
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Embodiment 1
[0035] Primers:
[0036]
[0037] 1. Preparation of AuNanostar@4-MBA@Au / sDNA SERS capture probe:
[0038] (1) Synthesis of gold species: Add 5 mL, 0.1 M triton and 5 mL, 0.5 mM chloroauric acid solution to a 25 mL round bottom flask, stir and mix at 300 r / min, then increase the speed to 800 r / min Add freshly prepared 0.6 mL, 0.01 M sodium borohydride in ice water, react for 2 min, and stand for 1 h to prepare the gold seed solution;
[0039] (2) Growth of AuNanostar: Add 10 mL, 0.1 M Triton, 10 mL, 0.5 mM chloroauric acid solution to a 25 mL round bottom flask, stir and mix at 300 r / min, then increase the speed to 800 r / min , add 12 μL, 0.5 mL, 1 mM silver nitrate and 0.6 mL, 0.0788 M ascorbic acid solution prepared in step (1) in sequence, react for 1 min, and get AuNanostar after standing for 1 h;
[0040] (3) Covalently bond 1mL, 2mM 4-MBA Raman signal molecules to the surface of the above-mentioned AuNanostar through the Au-S bond, and react at 400r / min for 30min to pre...
Embodiment 2
[0054] Embodiment 2 Detection limit calculation
[0055] According to the corresponding value of the SERS intensity of the blank experimental group in Table 2, the detection limit was calculated to be 0.25 fM.
[0056] Table 2 Blank group SERS corresponding value
[0057]
[0058] ;
[0059] y= 964.06 lgC OTA +5668.48;
[0060] 3 × S=A logC + B;
[0061] The calculated limit of detection was 0.25 fM.
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Abstract
The present invention discloses a method for detecting ochratoxin A based on SERS. The method comprises the following steps: fixing sDNA on surfaces of AuNanostatr @ 4-MBA @ Au core-shell starlike gold nanoparticles as an SERS capture probe and also fixing hairpin DNA on surfaces of magnetic beads. By utilizing high affinity of OTA and OTA aptamers and an Exo III cyclic amplification effect, a large amount of the AuNanostatr @ 4-MBA @ Au core-shell starlike gold nanoparticle SERS capture probes are connected to the surfaces of the magnetic beads. Therefore, intensity of the SERS obtained by detection has a close relation with concentration of the OTA, namely, an open mode of a SERS adapter sensor is developed for ultra-sensitive OTA detection. The provided detection method has characteristics of low cost, enhancement, high detection sensitivity, low detection line and good anti-interference performance for detecting the ochratoxin.
Description
technical field [0001] The invention belongs to the technical field of analysis and detection, and in particular relates to a method for detecting ochratoxin A based on SERS, which is applied in the field of toxin detection. Background technique [0002] Ochratoxin A (OTA), a mycotoxin that acts as a secondary metabolite of filamentous fungi such as Aspergillus and Penicillium, has attracted increasing attention as a food hazard. OTAs are commonly found in grains and other food during storage, because of their carcinogenic, teratogenic, mutagenic and immunosuppressive properties, so excessive intake of OTA may lead to acute or chronic disease episodes and even death. Therefore, there is an urgent need to develop a simple, sensitive method for the determination of OTA in contaminated food. [0003] In recent years, OTA detection technology has developed rapidly, including high performance liquid chromatography (HPLC), mass spectrometry (MS), HPLC-MS / MS, fluorescence, electro...
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