Nanoscale MoS2 has attracted extensive attention for sensing due to its superior properties. This study outlines a microfluidic and electrochemical biosensing methodology for the multiplex detection of paratuberculosis-specific miRNAs. Herein, we report the synthesis of MoS2 nanosheets decorated with a copper ferrite (CuFe2O4) nanoparticle composite and molecular probe immobilized MoS2 nanosheets as nanocarriers for the electrochemical detection of miRNAs. Paratuberculosis is a bacterial infection of the intestinal tract of dairy cattle, and is a cause of substantial economic and animal losses all over the world. The designed biosensing electrode was modified with the synthesized MoS2-CuFe2O4 nanocomposites for a highly amplified signal generation. Additionally, selective detection of miRNAs was accomplished by functionalizing the MoS2 nanosheets with a miRNA-specific biotin-tagged thiolated molecular probe and ferrocene thiol. The presence of target miRNA triggered the opening of the molecular probe present on the nanocarriers. The interaction of the molecular probe and miRNA resulted in an increase in the electrochemical signal from ferrocene. The optimized microfluidic biosensor was employed to detect a range of miRNA concentrations from the target analyte. Using square wave voltammetric analysis, a detection limit of 0.48 pM was calculated, with a detection range of 1 pM to 1.5 nM. The application of the biosensor was also assessed by detecting miRNAs in spiked serum and positive clinical samples. The developed nanomaterial enabled biosensor easily discriminated between the target miRNAs and other interfering molecules. The developed microfluidic biosensor has the potential to be used as a point-of-care, miRNA based diagnostic tool for paratuberculosis in dairy cows.
Fabrication of microsystems is traditionally achieved with photolithography. However, this fabrication technique can be expensive and non-ideal for integration with microfluidic systems. As such, graphene fabrication is explored as an alternative. This graphene fabrication can be achieved with graphite oxide undergoing optical exposure, using optical disc drives, to impose specified patterns and convert to graphene. This work characterises such a graphene fabrication, and provides fabrication, electrical, microfluidic, and scanning electron microscopy (SEM) characterisations. In the fabrication characterisation, a comparison is performed between traditional photolithography fabrication and the new graphene fabrication. (Graphene fabrication details are also provided.) Here, the minimum achievable feature size is identified and graphene fabrication is found to compare favourably with traditional photolithography fabrication. In the electrical characterisation, the resistivity of graphene is measured as a function of fabrication dose in the optical disc drive and saturation effects are noted. In the microfluidic characterisation, the wetting properties of graphene are shown through an investigation of the contact angle of a microdroplet positioned on a surface that is treated with varying fabrication dose. In the SEM characterisation, the observed effects in the previous characterisations are attributed to chemical or physical effects through measurement of SEM energy dispersive X-ray spectra and SEM images, respectively. Overall, graphene fabrication is revealed to be a viable option for development of microsystems and microfluidics.
Antibiotics are classes of antimicrobial substances that are administered widely in the field of veterinary science to promote animal health and feed efficiency. Cattle-administered antibiotics hold a risk of passing active residues to milk, during the milking process. This becomes a public health concern as these residues can cause severe allergic reactions to sensitive groups and considerable economic losses to the farmer. Hence, to ensure that the produced milk is safe to consume and adheres to permissible limits, an on-farm quick and reliable test is essential. This study illustrates the design and development of a microfluidic paper biosensor as a proof-of-concept detection system for gentamicin in milk. Localized surface plasmon resonance (LSPR) properties of gold nanoparticles have been explored to provide the user a visual feedback on the test, which was also corroborated by RGB analysis performed using Image J. The assay involves the use of a short stretch of single stranded DNA, called aptamer, which is very specific to the gentamicin present in the milk sample. The camera-based LOD for the fabricated paper device for milk samples spiked with gentamicin was calculated to be 300 nM, with a reaction time of 2 min.
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