West Nile Virus belongs to the RNA virus family of Flaviviridae. It is known to spread primarily through arthropod vectors such as mosquitoes. The virus infects mainly birds but is also reported to infect humans as well as pets such as cats and dogs. In humans, the majority of the infections caused by West Nile Virus are asymptomatic. However, some individuals (the majority of the confirmed cases) could enter a second, febrile stage with flu-like symptoms – commonly know as West Nile Fever. In a more serious stage, the disease becomes neuroinvasive, causing meningitis or encephalitis. Such severe conditions could lead to mortality. As the symptoms of West Nile Virus are very similar to other common diseases but can be fatal at a severe stage, it is important to distinguish the virus early in the diagnosis, particularly at the molecular level.
Norgen’s West Nile Virus RT-PCR Detection Kit is a ready-to-use system for the isolation and detection of WNV from blood or plasma samples. First, the kit contains components for the rapid isolation of total RNA, including viral RNA, from the samples using spin-column chromatography based on Norgen’s proprietary resin (Nucleic Acid Isolation). Second, the kit contains WNV Master Mix to allow for PCR Ampification, as well as a Control Master Mix to allow for amplification of both an isolation control and a PCR control. The amplified PCR products are then detected using agarose gel electrophoresis. Alternatively, detection can be performed based on real-time RT-PCR using melt curves (Detection). Please see the flowchart to the right.
The WNV RT-PCR Detection Mastermix contains reagents and enzymes for the specific amplification of a 356 bp region of the viral genome. In addition, Norgen’s WNV RT-PCR Detection Kit contains a second Mastermix, the RT-PCR Control Master Mix, which can be used to identify possible PCR inhibition and/or inadequate isolation via a separate RT-PCR reaction with the use of the provided PCR control (PCRC) or Isolation Control (IsoC), respectively. This kit is designed to allow for the testing of 24 samples. The WNV RT-PCR Primer Set and Controls are also available separately for end-point RT- PCR detection.
FEATURES AND BENEFITS
Isolate High Quality, Inhibitor-Free RNA - Removal of highly concentrated salts, metabolic wastes and proteins provides high quality, concentrated viral RNA.
Rapid Procedure - Isolate and detect West Nile Virus in under 3 hours.
Accuracy in Interpreting Results – Identify possible PCR inhibition and/or inadequate isolation with the provided RT-PCR Control Master Mix
High Sensitivity and Specificity - The limit of detection of the WNV2X RT-PCR Master Mix is 100 copies per reaction.
Purchase Primer Set and Controls Separately - The WNV Primer set as well as the positive control and negative control can be purchased separately