Electrochemical immunosensor for determination of Staphylococcus aureus bacteria by IgY immobilized on glassy carbon ele
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ORIGINAL PAPER
Electrochemical immunosensor for determination of Staphylococcus aureus bacteria by IgY immobilized on glassy carbon electrode with electrodeposited gold nanoparticles Mahmoud Roushani 1 & Zeinab Rahmati 1 & Mehdi Golchin 2 & Zinat Lotfi 2 & Mostafa Nemati 3 Received: 27 March 2020 / Accepted: 2 September 2020 # Springer-Verlag GmbH Austria, part of Springer Nature 2020
Abstract A new ultrasensitive immunosensor is proposed based on the covalently attached anti-protein A antibody (IgY) on deposited gold nanoparticle (AuNP)-modified glassy carbon electrode (GCE) for the electrochemical measurement of Staphylococcus aureus (S. aureus). Chicken IgY as a capture antibody provides highly selective and specific binding to the target bacteria and selectively captures the S. aureus in its three-dimensional space. Due to that it can eliminate the interference from protein G-producing Streptococcus. In addition, the electron-transfer characteristic of [Fe(CN)6]4−/3− is hindered by this combination; as it is reflected on the electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) curves. The proposed immunosensor displays a wide linear dynamic range from 10 to 107 CFU mL−1 with a detection limit of 3.3 CFU mL−1 with RSD 3.0%. It is capable to accurately determine S. aureus in milk and human blood serum as a complex matrix sample with satisfactory recovery of ∼ 97–103%. The immunosensor also displays high selectivity over other bacteria and acceptable stability. Presumably, our study can be regarded as the first one to report chicken IgY in order to detect S. aureus based on an electrochemical method. Keywords Staphylococcus aureus . Immunosensor . Anti-protein A antibody . Gold nanoparticles . Chicken IgY
Introduction S. aureus is known to be an important infectious agent worldwide [1–3]. One of the most important virulence factors of S. aureus is 42 kDa protein which is called surface immunoglobulin (Ig)-binding protein A (SpA) [4]. This protein binds to the Fc region of IgG antibodies and prevents disrupting the bacteria via opsonization and phagocytosis [5]. At present, clinical Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00604-020-04547-6) contains supplementary material, which is available to authorized users. * Mahmoud Roushani [email protected]; [email protected] 1
Department of Chemistry, Faculty of Sciences, Ilam University, PO Box 69315-516, Ilam, Iran
2
Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran
3
Department of Veterinary, Faculty of microbiology, Ilam University, PO Box 69315-516, Ilam, Iran
identification of pathogenic bacteria infection in bloodstream mainly depends on automated blood culture system. Generally, this method requires a long turnaround time of 24–72 h for the whole process [6], which is not conducive for rapid bacterial detection. Several strategies have been developed for the diagnosis, including enzyme-linked immunosorbent
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