Rapid and sensitive detection of box turtles using an electrochemical DNA biosensor based on a gold/graphene nanocomposite
Document Type
Article
Publication Date
12-6-2022
Abstract
The Southeast Asian box turtle, Cuora amboinensis, is an ecologically important endangered species which needs an onsite moni-toring device to protect it from extinction. An electrochemical DNA biosensor was developed to detect the C. amboinensis mito-chondrial cytochrome b gene based on an in silico designed probe using bioinformatics tools, and it was also validated in wet-lab experiments. As a detection platform, a screen-printed carbon electrode (SPCE) enhanced with a nanocomposite containing gold nanoparticles and graphene was used. The morphology of the nanoparticles was analysed by field-emission scanning electron microscopy and structural characteristics were analysed by using energy-dispersive X-ray, UV-vis, and Fourier-transform infrared spectroscopy. The electrochemical characteristics of the modified electrodes were studied by cyclic voltammetry, differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy. The thiol-modified synthetic DNA probe was immobilised on modified SPCEs to facilitate hybridisation with the reverse complementary DNA. The turtle DNA was distinguished based on hybridisation-induced electrochemical change in the presence of methylene blue compared to their mismatches, noncomplementary, and nontarget species DNA measured by DPV. The developed biosensor exhibited a selective response towards reverse comple-mentary DNAs and was able to discriminate turtles from other species. The modified electrode displayed good linearity for reverse complementary DNAs in the range of 1 x 10-11-5 x 10-6 M with a limit of detection of 0.85 x 10-12 M. This indicates that the pro-posed biosensor has the potential to be applied for the detection of real turtle species.
Keywords
Box turtle, DNA detection, Electrochemical DNA biosensor, Nanocomposite, Screen-printed carbon electrode
Divisions
InstituteofBiologicalSciences,nanocat
Funders
Nanotechnology and Catalysis Research Center, University of Malaya,Bangabandhu Science and Technolo-gy Fellowship Trust, Government of the People?s Republic of Bangladesh (Grant No: RU003-2021)
Publication Title
Beilstein Journal of Nanotechnology
Volume
13
Publisher
Beilstein-Institut
Publisher Location
TRAKEHNER STRASSE 7-9, FRANKFURT AM MAIN, 60487, GERMANY