Development of an aptamer/fluorescence dye PicoGreen-based method for detection of fumonisin B1
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National High Technology Research and Development Program of China (863 Program) (No. 2012AA101602), Project Supported by Zhejiang Province University Student Science and Technology Innovation Plan (Fresh Talent Program) (No. 2014R412040), Zhejiang A&F University Talent Starting Program (Nos. 2011FR025, 2013FR012, 2012FR047, 2013FR076, 2013FR054).

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    Abstract:

    Fumonisin B1 (FB1) is a carcinogenic mycotoxin found in commodities such as corn and corn-originated products. An aptamer-based method for detection of FB1 was developed using the fluorescent dye PicoGreen, which can recognize and bind double-stranded DNA. A peak fluorescence of PicoGreen was obtained in 15 min in the presence of FB1 aptamer, which formed a double-stranded hybridizer DNA with its complementary strand. The excitation and emission wavelengths for PicoGreen detection were 480 nm and 520 nm, respectively. The sensitivity of this aptamer/PicoGreen-based method was 0.1 μg/L. This method showed a good linearity for FB1 concentration ranging from 0.1 to 1 μg/L. The entire detection procedure for FB1 could be completed within 40 min. No cross reactions were observed with any other mycotoxins against aflatoxin B1, ochratoxin A, citrinin and zearalenone, demonstrating high specificity towards FB1 aptamer. Agreement between commercial, antibody-based enzyme-linked immunosorbent assay (ELISA) kit and aptamer method was excellent with a kappa value of 0.857. Taken together, this aptamer/PicoGreen-based method is more cost-effective, time-saving and useful than ELISA for detection of FB1.

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桂海娈,金庆日,张亚军,王晓杜,杨永春,邵春艳,程昌勇,卫芳芳,杨杨,杨梦华,宋厚辉. 基于荧光染料PicoGreen和核酸适配体的伏马毒素B1检测方法[J]. Chinese Journal of Biotechnology, 2015, 31(9): 1393-1400

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  • Received:November 30,2014
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  • Online: September 01,2015
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