Background

West Nile Virus (WNV) is an arbovirus that is transmitted through a complex life cycle alternating between invertebrate (i.e. mosquitoes) and vertebrate (i.e. birds) hosts. WNV infections can lead to fever, severe headaches, and body aches. In susceptible populations such as the elderly, the disease can progress to encephalitis, which can be fatal. Surveillance of mosquito populations for the incidence of WNV and measures to control the mosquito population are methods to mitigate the impact of WNV on public health.

Rapid and specific detection of West Nile Virus in environmental samples

Any verified laboratory method or commercial purification kits could be used to extract RNA from a variety of samples. The Loopamp® West Nile Virus Set used with the Loopamp® RNA Amplification Kit (sold separately) targets the envelope (env) gene of the genomic RNA. By using four primers to recognize six distinct regions in the gene, this test is highly specific. Due to the high amplification efficiency of LAMP, the results are achieved in 35 minutes.

Reverse transcription, amplification and detection in one single-tube with real-time results

In order to detect West Nile Virus using the LAMP method, the reverse-transcription and the cDNA amplification reactions are performed simultaneously in the same tube at 63°C. There is no extra step for cDNA synthesis.Reverse-transcription, amplification, and real-time detection are achieved in one step with Loopamp® Realtime Turbidimeters. Since electrophoresis is not required, the whole process is completed within one reaction tube, which greatly reduces the possibility of cross-contamination frequently associated with nucleic acid amplification technologies.

Alternative method of detection

Because RT-LAMP is an isothermal reaction, the testing can also be performed in a water bath or a heat block set at 63°C. The endpoint results can be visualized by adding Loopamp® Fluorescent Detection Reagent (sold separately) in the initial master mix.