Cholera is a potentially fatal diarrheal disease caused by Vibrio cholerae and is spread to humans from contaminated food and water. In order to prevent spread of epidemic chlorea, the development of novel sensitive, selective, user-friendly, cost-effective and rapid detection systems to detect of V. cholerae are necessary. Therefore, in this study, it was aimed to develop a specific, electrochemical immunoassay with high selectivitiy and sensitivity for detection of V. cholerae from drinking water using in house synthesized Gold Nanoparticles (AuNPs). The synthesized AuNPs were characterized by UV/Vis spectroscopy, Dynamic Light Scattering (DLS) and Atomic Force Microscopy (AFM) and electrochemical techniques were applied to confirm the succesful fabrication of the immunosensor. Also, this study focuses on the development of an antibody sensor for V. cholerae detection using a standard immunoassay without using nanoparticle. To accomplish that, in house spherical synthesized AuNPs at various sizes were synthesized, conjugated with secondary antibody-horseradish peroxidase enzyme (HRP) complex and their possible effect on the lowest detection limit of V. cholerae was investigated in comparison to commercially available AuNPs. The AuNPs-immunosensor on the results enabled the quantification of V. cholerae in a wide concentration range with a high sensitivity limit of detection (1 Colony-Forming Units/Milliliter) and specificity. Although the effect of 33 and 54 nm AuNPs on the process is close to each other, it has been observed that there is a 34% loss of efficiency when the size of the nanoparticle increases. With this study, a novel V. cholerae specific immunosensor was developed and the effects of in house synthesized AuNPs with various diameter on this developed biosensor were investigated in detail.
Keywords: AuNPs synthesis; Cholerae; Fast detection; Immunosensor; Rapid detection of bacteria; V.
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