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MICHÁLEK, P. DOSTÁLOVÁ, S. BUCHTELOVÁ, H. CERNEI, N. KREJČOVÁ, L. HYNEK, D. MILOSAVLJEVIĆ, V. JIMENEZ JIMENEZ, A. KOPEL, P. HEGER, Z. ADAM, V.
Original Title
A two-step protocol for isolation of influenza A (H7N7) virions and their RNA for PCR diagnostics based on modified paramagnetic particles
Type
journal article in Web of Science
Language
English
Original 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 (-Fe2O3) 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 - Fe2O3 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-ona- chip device. We assume that magnetic isolation is an exceptional procedure which can significantly accelerate the diagnostic possibilities of a broad spectrum of diseases.
Keywords
Influenza / Magnetic particles / PCR / Virions
Authors
MICHÁLEK, P.; DOSTÁLOVÁ, S.; BUCHTELOVÁ, H.; CERNEI, N.; KREJČOVÁ, L.; HYNEK, D.; MILOSAVLJEVIĆ, V.; JIMENEZ JIMENEZ, A.; KOPEL, P.; HEGER, Z.; ADAM, V.
Released
1. 7. 2016
ISBN
1522-2683
Periodical
Electrophoresis
Year of study
37
Number
14
State
Federal Republic of Germany
Pages from
2025
Pages to
2035
Pages count
11
BibTex
@article{BUT126809, author="Petr {Michálek} and Simona {Dostálová} and Hana {Michálková} and Natalia Vladimirovna {Cernei} and Ludmila {Krejčová} and David {Hynek} and Vedran {Milosavljević} and Ana Maria {Jimenez Jimenez} and Pavel {Kopel} and Zbyněk {Heger} and Vojtěch {Adam}", title="A two-step protocol for isolation of influenza A (H7N7) virions and their RNA for PCR diagnostics based on modified paramagnetic particles", journal="Electrophoresis", year="2016", volume="37", number="14", pages="2025--2035", doi="10.1002/elps.201600044", issn="1522-2683" }