title: A two-step protocol for isolation of influenza A (H7N7) virions and their RNA for PCR diagnostics based on modified paramagnetic particles.
abstract: Annual epidemics of influenza cause death of hundreds of thousands people and they also have a significant economic impact. Hence, a need for fast and cheap influenza diagnostic method is arising. The conventional methods for an isolation of the viruses are time-consuming and require expensive instrumentation as well as trained personnel. In this study, we modified the surface of nanomaghemite (γ-Fe2 O3 ) paramagnetic core with tetraethyl orthosilicate and (3-aminopropyl)triethoxysilane and the resulting particles were utilized for the isolation of H7N7 influenza virions. Consequently, we designed γ-Fe2 O3 paramagnetic core modified with calcium tripolyphosphate which was employed for the isolation of viral nucleic acid after virion's lysis. Both of these procedures can be performed rapidly in less than 10 min and, in combination with the RT-PCR, the whole influenza detection can be shortened to few hours. Moreover, the whole protocol could be easily automated and/or miniaturized, and thus can serve as a basis for use in a lab-on-a-chip device. We assume that magnetic isolation is an exceptional procedure which can significantly accelerate the diagnostic possibilities of a broad spectrum of diseases.
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University in Brno, Brno, Czech Republic.|Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Michalek
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University in Brno, Brno, Czech Republic.|Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
authors.forename: Simona
authors.initials: S
authors.lastname: Dostalova
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University in Brno, Brno, Czech Republic.
authors.forename: Hana
authors.initials: H
authors.lastname: Buchtelova
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University in Brno, Brno, Czech Republic.|Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
authors.forename: Natalia
authors.initials: N
authors.lastname: Cernei
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University in Brno, Brno, Czech Republic.|Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
authors.forename: Ludmila
authors.initials: L
authors.lastname: Krejcova
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University in Brno, Brno, Czech Republic.|Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
authors.forename: David
authors.initials: D
authors.lastname: Hynek
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University in Brno, Brno, Czech Republic.|Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
authors.forename: Vedran
authors.initials: V
authors.lastname: Milosavljevic
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University in Brno, Brno, Czech Republic.|Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
authors.forename: Ana Maria Jimenez
authors.initials: AM
authors.lastname: Jimenez
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University in Brno, Brno, Czech Republic.|Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
authors.forename: Pavel
authors.initials: P
authors.lastname: Kopel
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University in Brno, Brno, Czech Republic.|Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
authors.forename: Zbynek
authors.initials: Z
authors.lastname: Heger
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University in Brno, Brno, Czech Republic.|Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
authors.forename: Vojtech
authors.initials: V
authors.lastname: Adam
mesh_terms: D000818:Animals; D002642:Chick Embryo; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D053126:Influenza A Virus, H7N7 Subtype / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D048348:Reverse Transcription; D014771:Virion / Q000302:isolation & purification*
keywords: Influenza; Magnetic particles; PCR; Virions