Dispensable nucleic acid aptamer sensor and method for detecting okadaic acid by using sensor
A nucleic acid aptamer and sensor technology, applied in the field of sulfhydryl absorption, can solve the problems of difficulty in on-site, rapid detection, large amount of organic reagents, and difficult operation, and achieves simple and cheap synthesis and use, simple operation, and avoidance of Effects of Electrode Contamination
Inactive Publication Date: 2019-04-16
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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[0003] The technical problem solved by the present invention is to provide a disposable nucleic acid aptamer sensor and its detection method for okadaic acid, so as to provide a simple, quick, green and environmentally friendly technical means, which overcomes the expensive instruments and the lack of organic reagents in the existing detection methods. Large amount of use, difficult operation, difficult to achieve on-site, rapid detection and other defects
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[0025] The technical solution of the present invention will be further explained by the following examples, but the protection scope of the present invention is not limited in any form by the examples.
[0026] A disposable nucleic acid aptamer sensor and detection method thereof for highly sensitive detection of okadaic acid in shellfish, comprising the following steps:
[0027] (1) Sample pretreatment: Weigh 2 g of the homogenate sample, extract the toxin with 18 mL of methanol, vortex for 2 min, centrifuge at 8000 r / min for 5 min, and dilute the supernatant in a 20 mL volumetric flask with methanol. Accurately pipette 1 mL of the extract, add 125 μL of 2.5 mol / L NaOH solution, mix well, seal, and incubate at 76°C for 40 min. After the solution was cooled to room temperature, continue to add 125 μL of 2.5mol / L HCl solution to neutralize it, then blow it nearly dry with nitrogen, then add 1 mL of PBS buffer solution to redissolve, vortex and mix well, and then test.
[0028]...
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The invention discloses a dispensable nucleic acid aptamer sensor and a method for detecting okadaic acid by using the sensor and belongs to the technical field of aquatic product safety detection. According to the dispensable nucleic acid aptamer sensor and the method for detecting the okadaic acid by using the sensor of the invention, the nucleic acid aptamer of okadaic acid is used as a recognition probe; the recognition probe is fixed on the surface of the working electrode surface of a screen-printed gold electrode; electrochemical impedance spectroscopy is adopted to perform test; and the quantitatively detection of OA (okadaic acid) in shellfishes can be realized through a linear relationship between the change amount of the electron transfer impedance of the working electrode surface and OA concentration logarithmic values before and after the nucleic acid aptamer is combined with the okadaic acid. The sensor has the advantages of simplicity in preparation, great convenience and quickness, and low cost. The method for detecting the okadaic acid in the shellfishes by using the sensor is simple and highly efficient with a detection limit being 1 microgramme / kg, a linear rangebeing from 1 to 1800 microgramme / kg, a recovery rate ranging from 91.95% to 118.56%, and relative standard deviation ranging from 5.21% to 11.64%. With the method adopted, the screening and quantitative analysis of a large number of samples can be realized.
Description
technical field [0001] The invention relates to a throwable nucleic acid aptamer sensor and a method for detecting okadaic acid. Specifically, nucleic acid aptamers are used as identification probes, and core-shell nano-gold magnetic particles (Fe 3 o 4 @Au) and the sulfhydryl group modified at the 5' end of the nucleic acid aptamer immobilized the recognition probe on the surface of the working electrode of the screen-printed gold electrode (SPGE), and developed a disposable nucleic acid The aptamer sensor realizes the rapid and highly sensitive detection of okadaic acid in shellfish. The invention belongs to the technical field of aquatic product safety detection. Background technique [0002] Okadaic acid (OA) is the main toxic component of Diarrhetic shellfishpoisoning (DSP). Accumulates in digestive glands after filter-feeding intake. OA can cause nausea, vomiting, and diarrhea in those who eat it by mistake; it has a strong inhibitory effect on the activity of prot...
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IPC IPC(8): G01N27/327
CPCG01N27/3278
Inventor 郭萌萌谭志军陈佳琦吴海燕郑关超彭吉星翟毓秀
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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