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Updated: Mar 5, 2026
Vector Competence Analyses on Aedes aegypti Mosquitoes using Zika Virus
Published on: May 31, 2020
Rapid Surveillance for Vector Presence (RSVP): Development of a novel system for detecting Aedes aegypti and Aedes
Brian L Montgomery1, Martin A Shivas2, Sonja Hall-Mendelin3
1Metro South Public Health Unit, Queensland Health, Coopers Plains, Queensland, Australia.
Background:
The globally important Zika, dengue and chikungunya viruses are primarily transmitted by the invasive mosquitoes, Aedes aegypti and Aedes albopictus. In Australia, there is an increasing risk that these species may invade highly urbanized regions and trigger outbreaks. We describe the development of a Rapid Surveillance for Vector Presence (RSVP) system to expedite presence- absence surveys for both species.
Methodology/Principal Findings:
We developed a methodology that uses molecular assays to efficiently screen pooled ovitrap (egg trap) samples for traces of target species ribosomal RNA. Firstly, specific real-time reverse transcription-polymerase chain reaction (RT-PCR) assays were developed which detect a single Ae. aegypti or Ae. albopictus first instar larva in samples containing 4,999 and 999 non-target mosquitoes, respectively. ImageJ software was evaluated as an automated egg counting tool using ovitrap collections obtained from Brisbane, Australia. Qualitative assessment of ovistrips was required prior to automation because ImageJ did not differentiate between Aedes eggs and other objects or contaminants on 44.5% of ovistrips assessed, thus compromising the accuracy of egg counts. As a proof of concept, the RSVP was evaluated in Brisbane, Rockhampton and Goomeri, locations where Ae. aegypti is considered absent, present, and at the margin of its range, respectively. In Brisbane, Ae. aegypti was not detected in 25 pools formed from 477 ovitraps, comprising ≈ 54,300 eggs. In Rockhampton, Ae. aegypti was detected in 4/6 pools derived from 45 ovitraps, comprising ≈ 1,700 eggs. In Goomeri, Ae. aegypti was detected in 5/8 pools derived from 62 ovitraps, comprising ≈ 4,200 eggs.
Conclusions/Significance:
RSVP can rapidly detect nucleic acids from low numbers of target species within large samples of endemic species aggregated from multiple ovitraps. This screening capability facilitates deployment of ovitrap configurations of varying spatial scales, from a single residential block to entire suburbs or towns. RSVP is a powerful tool for surveillance of invasive Aedes spp., validation of species eradication and quality assurance for vector control operations implemented during disease outbreaks.
Insights
A new Rapid Surveillance for Vector Presence (RSVP) system uses molecular assays to detect invasive Aedes mosquitoes. This tool efficiently screens ovitrap samples, aiding in early detection and control of disease vectors.
Area of Science:
- Vector-borne diseases
- Invasive species ecology
- Molecular diagnostics
Background:
- Zika, dengue, and chikungunya viruses pose global health risks.
- Invasive Aedes aegypti and Aedes albopictus mosquitoes are primary vectors.
- Australia faces increasing risk of outbreaks in urbanized regions.
Purpose of the Study:
- To develop and evaluate a Rapid Surveillance for Vector Presence (RSVP) system.
- To expedite presence-absence surveys for Aedes aegypti and Aedes albopictus.
- To enhance early detection and control of invasive mosquito populations.
Main Methods:
- Development of real-time RT-PCR assays to detect target species ribosomal RNA.
- Screening of pooled ovitrap samples containing up to 4,999 non-target mosquitoes.
- Evaluation of ImageJ software for automated egg counting (with limitations).
- Field validation of the RSVP system in Brisbane, Rockhampton, and Goomeri.
Main Results:
- RSVP successfully detected Aedes aegypti in Rockhampton and Goomeri, where it is present or at the range margin.
- Aedes aegypti was not detected in ovitraps from Brisbane, where it is considered absent.
- The system efficiently screened large numbers of eggs using pooled samples.
Conclusions:
- RSVP enables rapid detection of invasive Aedes species using molecular assays.
- The system supports surveillance at various spatial scales, from blocks to towns.
- RSVP is a valuable tool for invasive species surveillance, eradication validation, and vector control quality assurance.
