The collection and quantification of influenza virus is important in determining exposure, and in the control of infection. There are a number of bioaerosol samplers and analytical methods available to do this, however few researchers have been able to detect influenza virus in the environment. Therefore, new samplers and methods should be considered.
This study compared influenza virus recovery from three filter types (PTFE, PVC, and PS) treated with sampling air or no air, using HBSS or PBS as wash buffers, and the QIAamp Viral RNA Mini Kit or Trizol method in the extraction of viral RNA. Twenty, 30-minute laboratory trials were completed in a sampling chamber designed to control RH at 50%. Samples were analyzed using RT-qPCR.
Viral recovery for each filter type was significantly different (p-value < .0001), with the new PS filter material resulting in the recovery of the most viral RNA. Treating filters with simulated sampling air did not affect the recovery of viral RNA from the filter materials (Kit, HBSS: Air vs. No Air p-value = 0.615; Trizol, HBSS: Air vs. No Air p-value = 0.947; Kit, PBS: Air vs. No Air p-value = 0.224; Trizol, PBS: Air vs. No Air p-value = 0.1122). Using HBSS and PBS as a filter wash buffer resulted in viral recoveries that were significantly different, depending on the RNA extraction method used (Kit, Air or No: HBSS vs. PBS p-value = 0.0001, 0.0001; Trizol, Air or No: HBSS vs. PBS p-value = 0.0322, 0.499). Viral RNA counts were significantly different when using the Kit and Trizol extraction methods for all comparisons (HBSS, Air or No-Air: Kit vs. Trizol p-value = 0.0021, 0.0013; PBS, Air or No-Air: Kit vs. Trizol p-value = 0.0001, 0.0002).
Our results demonstrated that the novel PS filter material resulted in the highest counts of extracted RNA compared to the commonly used PTFE and PVC, and that sampling air did not have a significant effect on viral recovery. Also, that the combination of HBSS with QIAamp Viral RNA Mini Kit, and PBS with the Trizol method, resulted in the highest counts of RNA extracted.
| Identifer | oai:union.ndltd.org:uiowa.edu/oai:ir.uiowa.edu:etd-7340 |
| Date | 01 August 2017 |
| Creators | Thedell, T. Aron |
| Contributors | Nonnenmann, Matthew W. |
| Publisher | University of Iowa |
| Source Sets | University of Iowa |
| Language | English |
| Detected Language | English |
| Type | thesis |
| Format | application/pdf |
| Source | Theses and Dissertations |
| Rights | Copyright © 2017 T. Aron Thedell |
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