Laboratory and Field Evaluations of a Commercially Available Real-Time Loop-Mediated Isothermal Amplification Assay for the Detection of West Nile Virus in Mosquito Pools.

Although the specific cDNA amplification mechanisms of reverse-transcriptase polymerase chain reaction (RT-PCR) and RT loop-mediated isothermal amplification (RT-LAMP) are very different, both molecular assays serve as options to detect arboviral RNA in mosquito pools. Like RT-PCR, RT-LAMP uses a reverse transcription step to synthesize complementary DNA (cDNA) from an RNA template and then uses target-specific primers to amplify cDNA to detectable levels in a single-tube reaction. Using laboratory-generated West Nile virus (WNV) samples and field-collected mosquito pools, we evaluated the sensitivity and specificity of a commercially available WNV real-time RT-LAMP assay (Pro-AmpRT™ WNV; Pro-Lab Diagnostics, Inc., Round Rock, Texas) and compared the results to a validated real-time RT-PCR assay. Laboratory generated virus stock samples containing ≥ 2.3 log10 plaque-forming units (PFU)/ml and intrathoracically inoculated mosquitoes containing ≥ 2.4 log10 PFU/ml produced positive results in the Pro-AmpRT WNV assay. Of field-collected pools that were WNV positive by real-time RT-PCR, 74.5% (70 of 94) were also positive by the Pro-AmpRT WNV assay, resulting in an overall Cohen's kappa agreement of 79.4% between the 2 tests. The Pro-AmpRT WNV assay shows promise as a suitable virus screening tool for vector surveillance programs provided agencies are aware of its characteristics and limitations.

Preparations and Activities Necessary for Aerial Mosquito Control After Hurricanes.

Vector Disease Control International (VDCI) has a long history of aiding mosquito control efforts necessary for recovery after natural disasters like hurricanes and major floods. As waters associated with these events begin to recede, both nuisance and vector mosquito species surge in abundance and consequently play an increased role in public health. When these situations arise, state and county agencies implement emergency response plans and many rely on Federal Emergency Management Agency or private contractors for assistance in reducing mosquito populations that can alter arbovirus transmission cycles, cause intolerable stress, hamper reconstruction efforts, and disrupt normal community functions. Vector Disease Control International owns the largest fleet of fixed-wing aircraft dedicated specifically to mosquito control and has worked every major storm event since Hurricane Bonnie in 1998. This article describes the logistics and operations required for implementing VDCI's emergency management plan, including the relocation of equipment, adult mosquito surveillance, delivery of pesticides, assessment of efficacy, and filing of low-level waivers and congested-area plans with the Federal Aviation Administration.