DNA Sensor for the Detection of Brucella spp. Based on Magnetic Nanoparticle Markers

Abstract

Due to the limitations of conventional Brucella detection methods, including safety concerns, long incubation times, and limited specificity, the development of 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 study, we introduce a magnetic nanoparticle marker-based biosensor using frequency mixing magnetic detection for point-of-care testing and quantification of Brucella DNA. Superparamagnetic nanoparticles were used as magnetically measured markers to selectively detect the target DNA hybridized with its complementary capture probes immobilized on a porous polyethylene filter. Experimental conditions like density and length of the probes, hybridization time and temperature, and magnetic binding specificity, sensitivity, and detection limit were investigated and optimized. Our sensor demonstrated a relatively fast detection time of approximately 10 min, with a detection limit of 55 copies (0.09 fM) when tested using DNA amplified from Brucella genetic material. 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. Our proposed biosensor has the potential to be used for the early detection of Brucella bacteria in the field and can contribute to disease control measures.

Nanomaterials-Integrated Electrochemical Biosensors as Pioneering Solutions for Zoonotic Disease Diagnosis

“…DNA sensor for the early identification of zoonotic pathogens. 94 Using the frequency mixing magnetic detection technique, we were able to quickly, selectively, and sensitively identify Brucella DNA on a functionalized PE filter by tapping into the non-linear and non-hysteretic magnetization properties of superparamagnetic nanoparticles. Reliable detection of Brucella DNA was achieved by the suggested magnetic biosensor, which was optimised and tested on synthetic and amplified gDNA.…”

Increasing the Particle Size and Magnetic Property of Iron Oxide Nanoparticles through a Segregated Nucleation and Growth Process

“…Therefore, they have a broad application prospect in the biomedical field. For in vivo applications, magnetic nanoparticles can not only be used as markers to assist in the detection and diagnosis of clinical pathology [11,12], but also as a tool for loading certain drugs to achieve directional transport and release [13,14]. For in vitro applications, the surface of magnetic nanoparticles can be modified with a variety of ligands (or receptors), which can be specifically bound to the receptors (or ligands) for the detection and separation of specific cells, viruses, proteins, and nucleic acids under the action of an applied magnetic field [15][16][17][18].…”