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A two-step protocol for isolation of influenza A (H7N7) virions and their RNA for PCR diagnostics based on modified paramagnetic particles
P. Michalek, S. Dostalova, H. Buchtelova, N. Cernei, L. Krejcova, D. Hynek, V. Milosavljevic, AM. Jimenez, P. Kopel, Z. Heger, V. Adam,
Jazyk angličtina Země Německo
Typ dokumentu časopisecké články
Odkazy
PubMed
27130152
DOI
10.1002/elps.201600044
Knihovny.cz E-zdroje
- MeSH
- chromatografie iontoměničová MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- kuřecí embryo MeSH
- polymerázová řetězová reakce metody MeSH
- reverzní transkripce MeSH
- virion izolace a purifikace MeSH
- virus chřipky A, podtyp H7N7 izolace a purifikace MeSH
- zvířata MeSH
- Check Tag
- kuřecí embryo MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
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.
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- $a Michalek, Petr $u Department of Chemistry and Biochemistry, Mendel University in Brno, Brno, Czech Republic. Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
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- $a A two-step protocol for isolation of influenza A (H7N7) virions and their RNA for PCR diagnostics based on modified paramagnetic particles / $c P. Michalek, S. Dostalova, H. Buchtelova, N. Cernei, L. Krejcova, D. Hynek, V. Milosavljevic, AM. Jimenez, P. Kopel, Z. Heger, V. Adam,
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- $a 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.
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- $a Dostalova, Simona $u Department of Chemistry and Biochemistry, Mendel University in Brno, Brno, Czech Republic. Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
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