Stability and detection of nucleic acid from viruses and hosts in controlled mosquito blood feeds

PLoS One. 2020 Jun 11;15(6):e0231061. doi: 10.1371/journal.pone.0231061. eCollection 2020.


Monitoring the presence and spread of pathogens in the environment is of critical importance. Rapid detection of infectious disease outbreaks and prediction of their spread can facilitate early responses of health agencies and reduce the severity of outbreaks. Current sampling methods are sorely limited by available personnel and throughput. For instance, xenosurveillance utilizes captured arthropod vectors, such as mosquitoes, as sampling tools to access blood from a wide variety of vertebrate hosts. Next generation sequencing (NGS) of nucleic acid from individual blooded mosquitoes can be used to identify mosquito and host species, and microorganisms including pathogens circulating within either host. However, there are practical challenges to collecting and processing mosquitoes for xenosurveillance, such as the rapid metabolization or decay of microorganisms within the mosquito midgut. This particularly affects pathogens that do not replicate in mosquitoes, preventing their detection by NGS or other methods. Accordingly, we performed a series of experiments to establish the windows of detection for DNA or RNA from human blood and/or viruses present in mosquito blood meals. Our results will contribute to the development of xenosurveillance techniques with respect to optimal timing of sample collection and NGS processing and will also aid trap design by demonstrating the stabilizing effect of temperature control on viral genome recovery from blood-fed mosquitoes.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blood*
  • Culicidae / virology*
  • DNA, Viral / analysis*
  • DNA, Viral / genetics
  • Environmental Monitoring
  • Humans
  • RNA, Viral / analysis*
  • RNA, Viral / genetics
  • Real-Time Polymerase Chain Reaction


  • DNA, Viral
  • RNA, Viral

Grants and funding

DEN and MEG were supported in part by the Hopkins Malaria Research Institute and Bloomberg Philanthropies, and MEG also received support from NIH 5T32AI007417. Commercial funder Microsoft Research provided financial support in the form of a gift to authors, James M. Pipas (JMP; University of Pittsburgh) and Douglas E. Norris (DEN; Johns Hopkins University). This gift was intended to support in the form of salaries for JMP and DEN. Authors Isaiah Hoyer, Andrzej Pastusiak, Michael R Reddy and Ethan K Jackson are paid employees of Microsoft Research. The funder provided support in the form of salaries for authors [IH, AP, MRR, EKJ], but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section.