Early Diagnosis of Leptospirosis Using Au Nanoparticle Sensor: nanosensor sensor ACS_AMI
By Dr. Priyom Bose, Ph.D.Aug 25 2022Reviewed by Megan Craig, M.Sc. Leptospirosis is a bacterial disease infecting both humans and animals. This zoonotic disease is most prevalent in tropical countries. In a recent ACS Applied Nano Materials study, researchers demonstrated the fabrication of an immunosensor that can rapidly and accurately detect leptospira in clinical samples.
What Do We Know About Leptospirosis? Among zoonotic diseases, leptospirosis is one of the leading causes of global morbidity and mortality, with approximately 1.03 million cases being reported every year. This disease is common in resource-limited populations, specifically in low to middle-income countries. The global impact of leptospirosis is not well understood because it is one of those diseases that has been neglected globally.
The available diagnostic methods for leptospirosis lack accuracy, sensitivity, portability, and repeatability. Mostly, for the clinical diagnosis of the disease, macroscopic agglutination test , conventional polymerase chain reaction , culture test, IgM enzyme-linked immunosorbent assay , high-resolution melting analysis, and real-time PCR are used.
AuNp rendered a significant enhancement of electron transfer rate and also assisted in the immobilization of specific molecules for the fabricated immunosensor. Glutathione was used to promote the formation of an elaborately ordered self-assembled monolayer on the AuNp, via the S−Au covalent bond. SAMs of GSH enabled the addition of a suitable amount of carboxylic acid as well as monoclonal antiLipL32 immobilization. The biological activity of the immobilized proteins was preserved by SAMs.
GSH concentration and the time required to form SAMs of GSH onto the AuNp-based electrode surface were optimized. Ten mM GSH was selected as the optimal concentration because it exhibited maximum redox potential due to the electrostatic interaction of the carboxyl group of GSH with the cationic probe. As the best oxidation current response was obtained at 10−6 g/mL antibody, this was selected to be the optimal antibody concentration for fabricating the new e-LipL32 sensor.
To further confirm the detection efficacy of the e-LipL32 sensor, the results of the tested clinical samples were compared with gold standard MAT, the IgM spot test, and IgM ELISA. Further, to differentiate the positive diagnosis of leptospirosis from other diseases, a boxplot was computed based on the averaged peak currents of all the clinical samples. A current above 9.8 μA was determined to be the threshold for positive leptospirosis diagnosis, which was confirmed by the MAT test.
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