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| Home > Publications database > Development of Superparamagnetic Based Biological Sensor for the Detection of Brucella DNA Using Frequency Mixing Magnetic Detection |
| Book/Dissertation / PhD Thesis | FZJ-2025-03256 |
2025
Forschungszentrum Jülich GmbH Zentralbibliothek, Verlag
Jülich
ISBN: 978-3-95806-836-0
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Please use a persistent id in citations: doi:10.34734/FZJ-2025-03256
Abstract: Early detection of zoonotic diseases is essential in preventing the consequences of outbreaks and reemergence occurrences. Brucellosis is endemic in several countries and has re-emerged with a high prevalence rate in different locations, affecting livestock and public health sectors. Due to the limitations of conventional Brucella detection methods, including limited specificity, long incubation times and safety concerns, developing a rapid, selective and accurate technique for the early detection of Brucella in livestock animals is crucial to prevent the spread of the associated disease. In the present thesis, we introduce a magnetic nanoparticle marker-based biosensor using Frequency Mixing Magnetic Detection (FMMD) for the detection and quantification of Brucella DNA. Magnetic nanoparticles (MNPs) were used as magnetically measured markers to selectively detect the target DNA hybridized with its complementary capture probes immobilized on a porous polyethylene filter. Our sensor demonstrated a relatively fast detection time of approximately 10 min, with a detection limit of 0.09 fM when tested using DNA amplified from Brucella genetic material by means of Polymerase Chain Reaction (PCR). In addition, the detection specificity was examined using gDNA from Brucella and other zoonotic bacteria that may coexist in the same niche, confirming the method’s selectivity for Brucella DNA. To enhance the practicality of the developed assay, we combined it with isothermal Recombinase Polymerase Amplification (RPA) and achieved rapid detection of 9 fM Brucella DNA in 25 minutes total assay time. In addition to isothermal DNA amplification in a water bath, we showed the feasibility of RPA directly inside our portable FMMD-based device. When being controlled by means of pulse width modulation (PWM), the inherently generated heat of the low frequency (LF) excitation coil of the magnetic reader can be utilized to serve as a constant temperature bath for RPA, thus enabling isothermal amplification inside the magnetic measurement head. We confirmed that RPA performs with high efficiency in the sensor unit of the FMMD device. In summary, the portability of the measurement device, the selective sensing of MNPs, the fast detection time, and the ability to deliver quantitative results make this biosensor a valuable tool for early on-site diagnosis and monitoring of Brucella infections in resource-limited settings.
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