Virus Detection Protocols for West Nile Virus in Vertebrate and Mosquito Specimens

• WNV belongs to the family Flaviviridae
• 2002 infecting 3,700 humans, in 44 states
• Detection of WNV looks for viral nucleic acid
• Other viruses found in mosquitoes, Flanders, Trivittatus, Cache Valley, and Highlands J
• RNeasy kits, used to screen RNA to find viral nucleic acid, done by hand
• Substitute for RNeasy kits is the ABI Prism 6700, work done my robotic workstation
• Some say that sample done by hand have less error in results, however ABI Prism 6700 showed the coeffients of variation, (accuracy) ranged from .2 to 5.5%, meaning it is very good at repeating RNA extraction with very little error
• RNA extraction from mosquito pools proved more difficult for the ABI Prism 6700
• To test accuracy, linearity, and range of detection, 50 mosquitoes were spiked with abdomens, heads, and appendages of positive WNV, all results agreed within 1 log
• Price differences of processing between RNeasy kits and ABI Prism 6700 is minimal
• Hands on time of ABI Prism 6700 is about one fourth of that of RNeasy kits.
• Robot method very accurate, with almost the same precision of that of human (RNeasy kit)
• Timesaving advantage may outweigh the initial cost of the robots