Method for detecting OTA by polymethacrylic acid LPFG

A technology of polymethacrylic acid and methacrylic acid, which is applied in the direction of measuring devices, biological testing, material inspection products, etc., can solve the problem of low response sensitivity, achieve the effect of enhancing response signal, reducing detection cost, and realizing high-sensitivity detection

Pending Publication Date: 2021-04-16
BIOLOGY INST OF SHANDONG ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In view of this, in order to solve the shortcoming that LPFG is not high to OTA solution response sensitivity, the present invention provides a kind of method that polymethacrylic acid LPFG detects OTA

Method used

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  • Method for detecting OTA by polymethacrylic acid LPFG

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Mix 0.78g of 2-methacrylic acid and 1.23g of polyethylene glycol dimethacrylate into 5ml of dimethyl sulfoxide solution, then add 0.3g of 2,2'-azobisisobutyronitrile, and stir well , pour the prepared solution into the liquid reaction pool for fixing LPFG, adjust the reaction temperature to 55°C, stop the reaction when the optimal sensitive range of the refractive index of LPFG is between 1.333-1.4000, and fully rinse with double distilled water, and use nitrogen gas blow dry. Then add EDC (0.04M) / NHS (0.01M) solution and let it stand for 1h to activate the carboxyl group to facilitate the connection of OTA monoclonal antibody. Then add 1 μM OTA monoclonal antibody and react overnight in the refrigerator at 4°C. Finally, OTA was detected by LPFG of monoclonal antibody modified with OTA.

Embodiment 2

[0041] Prepare 10ml of tetrahydrofuran solution with 0.07mol of 2-methacrylic acid and 0.03mol of polyethylene glycol dimethacrylate, stir well, then add 0.005mol of 2,2'-azobisisobutyronitrile, stir again After uniformity, pour the prepared solution into the liquid reaction pool where LPFG is fixed, adjust the reaction temperature to 65°C, stop the reaction when the optimal sensitive range of the refractive index of LPFG is between 1.333-1.4000, and fully rinse with twice distilled water , and blow dry with nitrogen. A solution of EDC (0.02M) / NHS (0.01M) was added and allowed to stand for 1 hour, then 1 μM OTA monoclonal antibody was added, and the mixture was reacted overnight in a refrigerator at 4°C. Finally, OTA was detected by LPFG of monoclonal antibody modified with OTA.

Embodiment 3

[0043] 5.36 grams of 2-methacrylic acid and 2.88 grams of polyethylene glycol dimethacrylate were prepared into 5ml of dimethyl sulfoxide solution, and then the two solutions were mixed, and then 0.33 grams of 2,2'-azobis Isobutyronitrile, stir to make it evenly mixed. Pour the prepared solution into the liquid reaction pool of fixed LPFG, adjust the temperature of the solution to 75°C, stop the reaction when the best sensitive range of the refractive index of LPFG is between 1.333-1.4000, and fully distilled water rinse. Add EDC (0.04M) / NHS (0.01M) solution and let stand for 1 hour, then add 10 μM OTA monoclonal antibody, and incubate at room temperature for 12 hours. Finally, OTA was detected by LPFG of monoclonal antibody modified with OTA.

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Abstract

The invention discloses a method for detecting OTA by polymethylacrylic acid LPFG, which comprises the following steps: continuously and controllably growing a polymethylacrylic acid film on the surface of LPFG in real time by controlling the concentration, reaction time and reaction temperature of a reaction reagent, so that the sensitivity of LPFG to the refractive index within the range of 1.333-1.4 reaches the optimal state. The OTA monoclonal antibody is further modified on the surface of the polymethacrylic acid film, the OTA detection selectivity of the LPFG is improved according to the specific reaction of the OTA monoclonal antibody and OTA, and the OTA content is judged according to the change of a resonance spectrum, so that the high-sensitivity and high-selectivity detection of OTA is realized. Fluorescence labeling is not needed, the detection steps are simplified, the detection cost is saved, the detection time is shortened, and the method is expected to be widely applied to the fields of food safety, food engineering, feed processing and the like.

Description

technical field [0001] The invention relates to the technical field of long-period grating detection, and more specifically relates to a method for detecting OTA with polymethacrylic acid LPFG. Background technique [0002] Ochratoxin A (OTA) is a very toxic mycotoxin, widely present in oats, barley, wheat, corn, animal feed and animal food (such as pig kidney, liver), etc. It is very harmful to health and can lead to renal failure, ureteral cancer, fatty liver, lymph node necrosis, and even death. Therefore, the detection of OTA is of great significance to the prevention of human diseases and the maintenance of people's health. [0003] Long period fiber grating (long period fiber grating, LPFG) is a passive optical sensing device sensitive to the refractive index of the environment. Many researchers have carried out research on LPFG in refractive index sensing, such as "Acta Optica Sinica" 2007,27 (010) published "CO 2 Research on the Refractive Index Characteristics of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/545G01N33/53
Inventor 李秋顺蔡雷马耀宏杨艳杨俊慧孟庆军王丙莲刘庆艾李大海
Owner BIOLOGY INST OF SHANDONG ACAD OF SCI
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