Abstract
Gene detection has important applications in biology, biomedical engineering, clinical, environmental, and marine fields. Rapid, sensitive, and selective recognition of specific genes is essential in practical applications. In this study, we describe a facile and sensitive DNA sensing platform for specific and quantitative detection of Heterosigma akashiwo, which is one of the causative agents of red tides. Fast and sensitive detection is achieved by using chemically synthesized graphene oxide (GO) nanosheets. Probe DNA is designed according to the specific DNA fragments of harmful algae and labeled with fluorescent molecules FAM (fluorescein-based dye). GO nanosheet solution is made, in which the strong interaction between FAM-labeled probe and GO nanosheets keeps them in close proximity, facilitating the fluorescence quenching of the fluorophore by GO nanosheets. In the presence of a complementary target DNA, the FAM-labeled DNA probe and the target DNA hybridize and desorb from the surface of GO nanosheets, resulting in restoration of fluorophore fluorescence. The concentration of target DNA fragments is analyzed by the fluorescence intensity at ~ 520 nm with emission wavelength of 480 nm. The sensitive detecting platform achieved stable measurement of 1 pM specific genes from Heterosigma akashiwo. Our GO nanosheet-based DNA-sensing platform performs fast and sensitive detection of trace amount of DNA, and enables quantitative recognition of harmful algae, which has promising applications in real-time monitoring in the marine environment of red tide generative dynamics, allowing effective control, particularly in relation to marine aquaculture.
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Funding
This work was financially supported by the National Key Research and Development Project of China (2019YFC1407800), Natural Science Foundation for Distinguished Young Scientist of Shandong Province (Grant No. JQ201814), Natural Science Foundation for Young Scientists of China (Grant No.61701282), Qilu Young Scholar Funds (11500086963060), The Fundamental Research Funds of Shandong University (Grant No. 2017JC020 and 2017TB0020), the National Natural Science Foundation of China (41876134), and Collaborative Innovation Center of Technology and Equipment for Biological Diagnosis and Therapy in Universities of Shandong.
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Le Qiang: Investigation, validation, formal analysis, writing—original draft; Yu Zhang: conceptualization, supervision; Chao Wu: resources; Yingkuan Han: validation; Suchun Wang: resources; Yanyan Wang: resources; Congcong Zhang: resources; Guangzhou Liu: resources; Qi Wu: resources; Hong Liu: project administration; Ian R. Jenkinson: supervision, writing; Jun Sun: supervision, project administration; Lin Han: supervision, methodology, resources, funding acquisition.
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Qiang, L., Zhang, Y., Wu, C. et al. A Facile and Sensitive DNA Sensing of Harmful Algal Blooms Based on Graphene Oxide Nanosheets. Mar Biotechnol 22, 498–510 (2020). https://doi.org/10.1007/s10126-020-09971-6
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DOI: https://doi.org/10.1007/s10126-020-09971-6