title: A luminescent hybridoma-based biosensor for rapid detection of V. cholerae upon induction of calcium signaling pathway.
abstract: In this study, a hybridoma based biosensor was developed for rapid, sensitive and selective detection of Vibrio cholerae O1 which converts the antibody-antigen binding to bioluminescence light. After investigation on hybridoma performance, the biosensor was constructed by transfecting specific hybridoma cells with aequorin reporter gene and the bioluminescence activities of stable biosensor were measured. The sensitivity of biosensor was as few as 50 CFU/ml and it showed no responses to other entric bacteria. Moreover, the response time of biosensor was estimated in 7th second which means this method is considerably faster than many available detection assays. In addition, this biosensor was successfully applied to V. cholerae detection in environmental samples with no significant loss in sensitivity, demonstrating our proposed biosensor provides a sensitive and reliable method for detection of V. cholerae in natural samples. The application of whole hybridoma cell directly as a sensing element in biosensor construction which mentioned for the first time in present study suggests that hybridoma cells could provide a valuable tool for future studies in both basic and diagnostic sciences and could be considered as a fast and specific sensing element for detection of other pathogens in different applications.
authors.affiliation: Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University,Tehran, Iran.
authors.forename: Parichehr
authors.initials: P
authors.lastname: Zamani
authors.affiliation: Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University,Tehran, Iran. Electronic address: sajedi_r@modares.ac.ir.
authors.forename: Reza H
authors.initials: RH
authors.lastname: Sajedi
authors.affiliation: Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University,Tehran, Iran.
authors.forename: Saman
authors.initials: S
authors.lastname: Hosseinkhani
authors.affiliation: Department of Bioscience and Biotechnology, Mallek Ashtar University of Technology, Tehran, Iran.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Zeinoddini
authors.affiliation: Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Bita
authors.initials: B
authors.lastname: Bakhshi
mesh_terms: D000937:Antigen-Antibody Reactions / Q000276:immunology; D015374:Biosensing Techniques; D020013:Calcium Signaling; D002771:Cholera / Q000175:diagnosis* / Q000382:microbiology; D006801:Humans; D006825:Hybridomas / Q000737:chemistry / Q000276:immunology; D049449:Luminescence; D014734:Vibrio cholerae / Q000302:isolation & purification* / Q000378:metabolism; D041241:Vibrio cholerae O1
keywords: Aequorin; Biosensor; CANARY technology; Hybridoma cell; Stable cell line; Vibrio cholerae