PMID: 11931239Apr 5, 2002Paper

Effect of holding conditions on the detection of West Nile viral RNA by reverse transcriptase-polymerase chain reaction from mosquito (Diptera: Culicidae) pools

Journal of Medical Entomology
Michael J TurellCharles E Cannon

Abstract

We evaluated the effect of holding temperature and time between mosquito death and processing mosquito pools for virus detection on our ability to detect West Nile (WN) viral RNA from pools of mosquitoes by reverse transcriptase-polymerase chain reaction (RT-PCR). Pools of 24 uninfected Culex pipiens L. mosquitoes were "spiked" with either a single Cx. pipiens that had been inoculated previously with WN virus or with an uninfected mosquito. These pools were held dry at 20, 4, -20, or -70 degrees C for selected time intervals before all mosquito pools were triturated in TRIzol LS reagent and processed for detection of WN viral RNA. While infectious virus virtually disappeared from pools maintained at 20 degrees C by 48 h after mosquito death, neither holding temperature (20 to -70 degrees C) nor holding period (up to 2 wk) affected detection of WN viral RNA by real-time RT-PCR. These findings suggest that we need not keep mosquitoes chilled to be able to detect WN viral RNA effectively by RT-PCR. This should enhance the feasibility of field-based WN virus surveillance programs where only detection of WN viral RNA is the objective and maintenance of a cold chain may not be possible.

References

Nov 1, 1974·The American Journal of Tropical Medicine and Hygiene·L Rosen, D Gubler
Oct 19, 2000·The American Journal of Tropical Medicine and Hygiene·M J TurellJ Oliver
May 16, 2001·Molecular and Cellular Probes·T L HadfieldE J Park

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Citations

Jan 9, 2003·Journal of Clinical Microbiology·Amy J LambertRobert S Lanciotti
Jan 11, 2012·The American Journal of Tropical Medicine and Hygiene·Mangala MavaleAkhilesh Chandra Mishra
Sep 23, 2011·Journal of Medical Entomology·Ting-Wu ChuangMark L Wilson
Dec 8, 2009·Journal of Medical Entomology·Jimmy B PitzerDonald F Caccamise
Apr 5, 2008·Journal of Virological Methods·Jamie A BlowDavid J Dohm
Apr 17, 2012·Journal of Biomedicine & Biotechnology·Andrew F van den HurkScott A Ritchie
Jul 1, 2016·Parasites & Vectors·Mohamed F SallamPhillip G Koehler
Jan 12, 2017·Emerging Infectious Diseases·Kristen L Burkhalter, Harry M Savage
Jun 1, 2007·Journal of the American Mosquito Control Association·B G BollingB J Beaty
Oct 28, 2006·Journal of the American Mosquito Control Association·Richard LampmanRobert Novak
May 14, 2017·The American Journal of Tropical Medicine and Hygiene·Sonja Hall-MendelinAndrew F van den Hurk
Apr 23, 2016·Journal of the American Mosquito Control Association·Elizabeth S Andrews, Michael J Turell
Jul 28, 2018·Journal of Medical Entomology·Daniel R TimminsScott A Ritchie
Jan 12, 2022·PLoS Neglected Tropical Diseases·UNKNOWN PLOS Neglected Tropical Diseases Editors

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