Automated, simple, and efficient influenza RNA extraction from clinical respiratory swabs using TruTip and epMotion

Abstract Background Rapid, simple and efficient influenza RNA purification from clinical samples is essential for sensitive molecular detection of influenza infection. Automation of the TruTip extraction method can increase sample throughput while maintaining performance. Objectives To automate TruT...

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Veröffentlicht in:Journal of clinical virology 2013-09, Vol.58 (1), p.138-143
Hauptverfasser: Griesemer, Sara B, Holmberg, Rebecca, Cooney, Christopher G, Thakore, Nitu, Gindlesperger, Alissa, Knickerbocker, Christopher, Chandler, Darrell P, St. George, Kirsten
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Sprache:eng
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Zusammenfassung:Abstract Background Rapid, simple and efficient influenza RNA purification from clinical samples is essential for sensitive molecular detection of influenza infection. Automation of the TruTip extraction method can increase sample throughput while maintaining performance. Objectives To automate TruTip influenza RNA extraction using an Eppendorf epMotion robotic liquid handler, and to compare its performance to the bioMerieux easyMAG and Qiagen QIAcube instruments. Study design Extraction efficacy and reproducibility of the automated TruTip/epMotion protocol was assessed from influenza-negative respiratory samples spiked with influenza A and B viruses. Clinical extraction performance from 170 influenza A and B-positive respiratory swabs was also evaluated and compared using influenza A and B real-time RT-PCR assays. Results TruTip/epMotion extraction efficacy was 100% in influenza virus-spiked samples with at least 745 influenza A and 370 influenza B input gene copies per extraction, and exhibited high reproducibility over four log10 concentrations of virus ( 0.90). Conclusion The automated TruTip/epMotion protocol is a simple and rapid extraction method that reproducibly purifies influenza RNA from respiratory swabs, with comparable efficacy and efficiency to both the easyMAG and QIAcube instruments.
ISSN:1386-6532
1873-5967
DOI:10.1016/j.jcv.2013.06.033