title: Piezoelectric immunochip coated with thin films of bacterial cellulose nanocrystals for dengue detection.
abstract: Low-cost piezoelectric devices, such as simple frequency monitoring quartz crystal microbalance (QCM) devices, have good clinical utility as fast diagnostic tools for the detection of several diseases. However, unspecific antigen recognition, poor molecular probe adsorption and the need for sample dilution are still common drawbacks that hinder their use in routine diagnosis. In this work, piezoelectric sensors were previously coated with thin films of bacterial cellulose nanocrystals (CN) to provide a more sensitive and adapted interface for the attachment of monoclonal immunoglobulin G (IgGNS1) and to favor specific detection of non-structural protein 1 (NS1) of dengue fever. The assembly of the immunochip surface was analyzed by atomic force microscopy (AFM) and the NS1 detection was followed by quartz crystal microbalance with (QCM-D) and without energy dissipation monitoring (QCM). The CN surface was able to immobilize 2.30±0.5mgm-2of IgGNS1, as confirmed by AFM topography and phase images along with QCM-D. The system was able to detect the NS1 protein in serum with only 10-fold dilution in the range of 0.01-10µgmL-1by both QCM and QCM-D. The limits of detection of the two devices were 0.1μgmL-1for QCM-D and 0.32μgmL-1for QCM. As a result, QCM-D and QCM apparatuses can be used to follow NS1 recognition and have good potential for more sensitive, fast and/or less expensive diagnostic assays for dengue.
authors.affiliation: Biochemistry and Molecular Biology Department, Federal University of Paraná, Centro Politécnico, CEP 81531-980 Curitiba, PR, Brazil; BioPol, Chemistry Department, Federal University of Paraná, P.B 19032, Centro Politécnico, CEP 81531-980 Curitiba, PR, Brazil.
authors.forename: Cleverton Luiz
authors.initials: CL
authors.lastname: Pirich
authors.affiliation: BioPol, Chemistry Department, Federal University of Paraná, P.B 19032, Centro Politécnico, CEP 81531-980 Curitiba, PR, Brazil.
authors.forename: Rilton Alves
authors.initials: RA
authors.lastname: de Freitas
authors.affiliation: Departamento de Química Fundamental, Instituto de Química, Universidade de São Paulo (USP), CP 26077, 05513-970 São Paulo-SP, Brazil.
authors.forename: Roberto Manuel
authors.initials: RM
authors.lastname: Torresi
authors.affiliation: BioPol, Chemistry Department, Federal University of Paraná, P.B 19032, Centro Politécnico, CEP 81531-980 Curitiba, PR, Brazil.
authors.forename: Guilherme Fadel
authors.initials: GF
authors.lastname: Picheth
authors.affiliation: Biochemistry and Molecular Biology Department, Federal University of Paraná, Centro Politécnico, CEP 81531-980 Curitiba, PR, Brazil; BioPol, Chemistry Department, Federal University of Paraná, P.B 19032, Centro Politécnico, CEP 81531-980 Curitiba, PR, Brazil. Electronic address: mariarita.sierakowski@gmail.com.
authors.forename: Maria Rita
authors.initials: MR
authors.lastname: Sierakowski
mesh_terms: D055910:Antibodies, Immobilized / Q000737:chemistry; D001419:Bacteria / Q000737:chemistry; D015374:Biosensing Techniques / Q000295:instrumentation; D002482:Cellulose / Q000737:chemistry; D003715:Dengue / Q000097:blood / Q000175:diagnosis* / Q000821:virology; D003716:Dengue Virus / Q000302:isolation & purification*; D004867:Equipment Design; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation; D056656:Lab-On-A-Chip Devices; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D011135:Polysaccharides, Bacterial / Q000737:chemistry*; D058525:Quartz Crystal Microbalance Techniques / Q000295:instrumentation*; D017361:Viral Nonstructural Proteins / Q000032:analysis / Q000097:blood*
chemical_list: D055910:Antibodies, Immobilized; D011135:Polysaccharides, Bacterial; D017361:Viral Nonstructural Proteins; D002482:Cellulose
keywords: Biosensors; Cellulose nanocrystals; Dengue; NS1 protein; Quartz crystal microbalance