Use of Centrifugal Filter Devices to Concentrate Dengue Virus in Mosquito per os Infection Experiments

PLoS One. 2015 Sep 15;10(9):e0138161. doi: 10.1371/journal.pone.0138161. eCollection 2015.

Abstract

Dengue virus (DENV) is an arbovirus transmitted to humans by the bite of infected Aedes mosquitoes. Experimental per os infection of mosquitoes with DENV is usually a preliminary step in virus/vector studies but it requires being able to prepare artificial blood-meals with high virus titers. We report here the convenient use of centrifugal filter devices to quickly concentrate DENV particles in cell-culture supernatants. The median viral titer in concentrated-supernatants was 8.50 log10 TCID50/mL. By using these DENV concentrated-supernatants to prepare infectious blood-meals in Aedes aegypti per os infection experiments, we obtained a mean mosquito-infection rate of 94%. We also evaluated the use of centrifugal filter devices to recover DENV particles from non-infectious blood-meals presented to infected mosquitoes through a feeding membrane to collect their saliva.

MeSH terms

  • Aedes / virology*
  • Animals
  • Blood / virology
  • Centrifugation / instrumentation*
  • Dengue / virology*
  • Dengue Virus / isolation & purification*
  • Dengue Virus / physiology
  • Filtration / instrumentation*
  • Insect Vectors / virology
  • Saliva / virology

Grants and funding

The authors have no support or funding to report.