Field-deployable molecular diagnostic platform for arbovirus detection in Aedes aegypti
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(2020) 13:489 Rutkowski et al. Parasites Vectors https://doi.org/10.1186/s13071-020-04357-y
Open Access
RESEARCH
Field‑deployable molecular diagnostic platform for arbovirus detection in Aedes aegypti Natalie Rutkowski, Yuemei Dong and George Dimopoulos*
Abstract Background: Surveillance of mosquito infection status is critical for planning and deployment of proper mosquito control initiatives. Point-of-care (POC) detection assays are necessary for monitoring the infection prevalence and geographical range of viruses in mosquito vector populations. We therefore assessed the novel real-time PCR (qPCR) bCUBE (Hyris, London, UK) molecular diagnostic system as a tool for virus detection. Methods: Aedes aegypti Rps17 was used to validate and determine correlation coefficient for the novel bCUBE qPCR system to a laboratory standard StepOnePlus real-time PCR system (Applied Biosystems, Waltham, MA, USA). Experimentally infected Ae. aegypti were quantified for Zika (ZIKV) and dengue virus serotype 2 (DENV2) viral genomic RNA. Infection prevalence was compared to plaque assay. Results: We developed and validated a novel qPCR system for the detection of ZIKV and DENV2 using the real-time qPCR system bCUBE. With bCUBE-based qRT-PCR, viral genomic RNA could be detected in individually infected Ae. aegypti mosquitoes and in pools of 5, 10 or 15 mosquitoes. Conclusions: The portable qPCR bCUBE diagnostic system is capable of detecting Zika and dengue virus in mosquitoes and therefore has potential as a practical field-deployable diagnostic test for vector-borne disease surveillance programmes. Keywords: Aedes aegypti, Zika virus, Dengue virus, Diagnostics, qPCR
Background Arthropod-borne diseases threaten over two-thirds of the global population and are exhibiting an ongoing expansion of their geographical range and prevalence as a result of climate change, urbanization and globalization [1–4]. According to the World Health Organization, dengue virus infection has increased 30-fold over the past 50 years and now affects nearly 100 million people, primarily in the Americas and Asia [5, 6]. The outbreaks of *Correspondence: [email protected] W. Harry Feinstone Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD, USA
Zika in 2015 and 2016 have also led to widespread concern because of the virus ability to cause newborn malformations [7, 8]. Dengue and Zika viruses are members of the genus Flavivirus that are primarily transmitted by the Aedes mosquitoes [7, 9, 10]. Other members of the genus Flavivirus are also recognized as vector-borne pathogens of public health significance, including West Nile virus (WNV), yellow fever virus (YFV), Japanese encephalitis virus (JEV) and chikungunya virus (CHIKV). These viruses cause similar flu-like symptoms with the potential to progress to neuroinvasive outcomes. Aedes mosquitoes, with their aggressive blood-feeding behavior, have allowed for efficient human-mosquito transmission of
© The Author(s) 2020.
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