title: An Elegant Analysis of White Spot Syndrome Virus Using a Graphene Oxide/Methylene Blue based Electrochemical Immunosensor Platform.
abstract: White spot syndrome virus (WSSV) is a major devastating virus in aquaculture industry. A sensitive and selective diagnostic method for WSSV is a pressing need for the early detection and protection of the aquaculture farms. Herein, we first report, a simple electrochemical immunosensor based on methylene blue dye (MB) immobilized graphene oxide modified glassy carbon electrode (GCE/GO@MB) for selective, quick (35 ± 5 mins) and raw sample analysis of WSSV. The immunosensor was prepared by sequential modification of primary antibody, blocking agent (bovine serum album), antigen (as vp28 protein), secondary antibody coupled with horseradish peroxidase (Ab2-HRP) on the GCE/GO@MB. The modified electrode showed a well-defined redox peak at an equilibrium potential (E1/2), -0.4 V vs Ag/AgCl and mediated H2O2reduction reaction without any false positive result and dissolved oxygen interferences in pH 7 phosphate buffer solution. Under an optimal condition, constructed calibration plot was linear in a range of 1.36 × 10-3to 1.36 × 107copies μL-1of vp28. It is about four orders higher sensitive than that of the values observed with polymerase chain reaction (PCR) and western blot based WSSV detection techniques. Direct electrochemical immunosensing of WSSV in raw tissue samples were successfully demonstrated as a real sample system.
authors.affiliation: Aquaculture Biotechnology Laboratory, Department of Integrative Biology, School of Biosciences and Technology, Vellore Institute of Technology University, Vellore-632014, Tamil Nadu, India.
authors.forename: Anusha
authors.initials: A
authors.lastname: Natarajan
authors.affiliation: Nano and Bioelectrochemistry Research Laboratory, Department of Chemistry, School of Advanced Sciences, Vellore Institute of Technology University, Vellore-632014, Tamil Nadu, India.
authors.forename: K S Shalini
authors.initials: KS
authors.lastname: Devi
authors.affiliation: Aquaculture Biotechnology Laboratory, Department of Integrative Biology, School of Biosciences and Technology, Vellore Institute of Technology University, Vellore-632014, Tamil Nadu, India.
authors.forename: Sudhakaran
authors.initials: S
authors.lastname: Raja
authors.affiliation: Nano and Bioelectrochemistry Research Laboratory, Department of Chemistry, School of Advanced Sciences, Vellore Institute of Technology University, Vellore-632014, Tamil Nadu, India.|Carbon dioxide Research and Green Technology Centre, Vellore Institute of Technology University, Vellore 632014, Tamil Nadu, India.
authors.forename: Annamalai
authors.initials: A
authors.lastname: Senthil Kumar
mesh_terms: D000818:Animals; D000906:Antibodies / Q000378:metabolism; D000941:Antigens / Q000378:metabolism; D015374:Biosensing Techniques / Q000379:methods*; D003386:Brachyura / Q000821:virology; D055664:Electrochemical Techniques / Q000379:methods*; D004566:Electrodes; D005880:Gills / Q000821:virology; D005898:Glass / Q000737:chemistry; D006108:Graphite / Q000737:chemistry*; D006735:Horseradish Peroxidase / Q000378:metabolism; D007118:Immunoassay / Q000379:methods*; D008751:Methylene Blue / Q000737:chemistry*; D010087:Oxides / Q000737:chemistry*; D046848:White spot syndrome virus 1 / Q000302:isolation & purification*
chemical_list: D000906:Antibodies; D000941:Antigens; D010087:Oxides; D006108:Graphite; D006735:Horseradish Peroxidase; D008751:Methylene Blue