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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

tetano

Editor, Senior Moderator
Electrophoresis. 2016 Apr 30. doi: 10.1002/elps.201600044. [Epub ahead of print]
[h=1]A two-step protocol for isolation of influenza A (H7N7) virions and their RNA for PCR diagnostics based on modified paramagnetic particles.[/h] Michalek P[SUP]1,[/SUP][SUP]2[/SUP], Dostalova S[SUP]1,[/SUP][SUP]2[/SUP], Buchtelova H[SUP]1[/SUP], Cernei N[SUP]1,[/SUP][SUP]2[/SUP], Krejcova L[SUP]1,[/SUP][SUP]2[/SUP], Hynek D[SUP]1,[/SUP][SUP]2[/SUP], Milosavljevic V[SUP]1,[/SUP][SUP]2[/SUP], Jimenez AM[SUP]1,[/SUP][SUP]2[/SUP], Kopel P[SUP]1,[/SUP][SUP]2[/SUP], Heger Z[SUP]1,[/SUP][SUP]2[/SUP], Adam V[SUP]1,[/SUP][SUP]2[/SUP].
[h=3]Author information[/h]

[h=3]Abstract[/h] 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-Fe[SUB]2[/SUB] O[SUB]3[/SUB] ) paramagnetic core with tetraethyl orthosilicate and (3-aminopropyl)triethoxysilane and the resulting particles were utilized for isolation of H7N7 influenza virions. Consequently, we designed γ-Fe[SUB]2[/SUB] O[SUB]3[/SUB] 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 minutes and, in the combination with RT-PCR, the whole influenza detection can be shortened to few hours. Moreover, the whole protocol can be easily automated and/or miniaturized, and thus can serve as a basis for use in a lab-on-a-chip device. We postulate that magnetic isolation is an exceptional procedure, which can significantly accelerate the diagnostic possibilities of a broad spectrum of diseases. This article is protected by copyright. All rights reserved.
This article is protected by copyright. All rights reserved.


[h=4]KEYWORDS:[/h] Influenza; Magnetic particles; PCR; Virions

PMID: 27130152 [PubMed - as supplied by publisher]
 
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