title: Biosensor for detection of antibiotic resistant Staphylococcus bacteria.
abstract: A structurally transformed lytic bacteriophage having a broad host range of Staphylococcus aureus strains and a penicillin-binding protein (PBP 2a) antibody conjugated latex beads have been utilized to create a biosensor designed for discrimination of methicillin resistant (MRSA) and sensitive (MSSA) S. aureus species (1,2). The lytic phages have been converted into phage spheroids by contact with water-chloroform interface. Phage spheroid monolayers have been moved onto a biosensor surface by Langmuir-Blodgett (LB) technique (3). The created biosensors have been examined by a quartz crystal microbalance with dissipation tracking (QCM-D) to evaluate bacteria-phage interactions. Bacteria-spheroid interactions led to reduced resonance frequency and a rise in dissipation energy for both MRSA and MSSA strains. After the bacterial binding, these sensors have been further exposed to the penicillin-binding protein antibody latex beads. Sensors analyzed with MRSA responded to PBP 2a antibody beads; although sensors inspected with MSSA gave no response. This experimental distinction determines an unambiguous discrimination between methicillin resistant and sensitive S. aureus strains. Equally bound and unbound bacteriophages suppress bacterial growth on surfaces and in water suspensions. Once lytic phages are changed into spheroids, they retain their strong lytic activity and show high bacterial capture capability. The phage and phage spheroids can be utilized for testing and sterilization of antibiotic resistant microorganisms. Other applications may include use in bacteriophage therapy and antimicrobial surfaces.
authors.affiliation: Department of Anatomy, Physiology and Pharmacology, College of Veterinary Medicine, Auburn University, USA.
authors.forename: Rajesh
authors.initials: R
authors.lastname: Guntupalli
authors.forename: Iryna
authors.initials: I
authors.lastname: Sorokulova
authors.forename: Eric
authors.initials: E
authors.lastname: Olsen
authors.forename: Ludmila
authors.initials: L
authors.lastname: Globa
authors.forename: Oleg
authors.initials: O
authors.lastname: Pustovyy
authors.forename: Vitaly
authors.initials: V
authors.lastname: Vodyanoy
mesh_terms: D001435:Bacteriophages / Q000502:physiology*; D015374:Biosensing Techniques / Q000379:methods*; D018796:Immunoconjugates / Q000737:chemistry; D055624:Methicillin-Resistant Staphylococcus aureus / Q000145:classification* / Q000821:virology; D008863:Microspheres; D046915:Penicillin-Binding Proteins / Q000737:chemistry; D010453:Peptide Synthases / Q000737:chemistry; D058525:Quartz Crystal Microbalance Techniques / Q000295:instrumentation / Q000379:methods; D013211:Staphylococcus aureus / Q000145:classification* / Q000821:virology
chemical_list: D018796:Immunoconjugates; D046915:Penicillin-Binding Proteins; C478699:penicillin-binding protein 2a, Streptococcus; D010453:Peptide Synthases