A new high-throughput tool to screen mosquito-borne viruses in Zika virus endemic/epidemic areas

Abstract

44Mosquitoes are vectors of arboviruses affecting animal and human health. Arboviruses circulate 45 primarily within an enzootic cycle and recurrent spillovers contribute to the emergence of 46 human-adapted viruses able to initiate an urban cycle involving anthropophilic mosquitoes. The 47 increasing volume of travel and trade offers multiple opportunities for arbovirus introduction 48 in new regions. This scenario has been exemplified recently with the Zika pandemic. To 49 incriminate a mosquito as vector of a pathogen, several criteria are required such as the 50 detection of natural infections in mosquitoes. In this study, we used a high-throughput chip 51 based on the BioMark TM Dynamic arrays system capable of detecting 64 arboviruses in a single 52 experiment. A total of 17,958 mosquitoes collected in Zika-endemic/epidemic countries 53 (Brazil, French Guiana, Guadeloupe, Suriname, Senegal, and Cambodia) were analyzed. Here 54 we show that this new tool can detect endemic and epidemic viruses in different mosquito 55 species in an epidemic context. Thus, this fast and low-cost method can be suggested as a novel 56 epidemiological surveillance tool to identify circulating arboviruses. 57 58 Other arboviruses such as West Nile virus (WNV; Flavivirus, Flaviviridae) remain circulating 84within an enzootic cycle with sporadic spillovers causing human cases. 85Many regions experience simultaneous circulation of different arboviruses (6, 7), and co-86 infections in vectors were reported (8). These coinfections can present an opportunity for 87 viruses to exchange genetic material. Impacts of such genetic events on virulence for vertebrate 88 hosts are still unknown (9). Thus, being able to detect a wide range of arboviruses in thousands 89 of field-collected mosquitoes in a single experiment can be a valuable tool to predict arboviral 90 emergences in human populations. Indeed similar methods were developed with success to 91 screen tick-borne pathogens (bacteria, parasites and viruses) and allowed the detection of 92 expected and unexpected pathogens in large scale epidemiological studies (10, 11). Therefore, 93 we developed a high throughput system based on real-time microfluidic PCR which is able to 94 detect 96 mosquito-borne viruses in 96 samples within one single run. With this method, we 95 have screened: (1) mosquitoes infected artificially using a feeding system to validate our tool, 96(2) mosquitoes collected in countries endemic for the major human arboviruses (e.g., Senegal, 97 Cambodia, Brazil), and (3) mosquitoes collected during the Zika and Yellow fever outbreaks 98 in the Americas (French Guiana, Guadeloupe, Brazil, Suriname). This method allowed 99 detecting epidemic viruses (ZIKV, CHIKV, YFV) but also unexpected viruses (e.g. Trivittatus 100 virus, TVTV, Orthobunyavirus, Bunyaviridae) underlining the need of such a tool for early 101 detection of emerging mosquito-borne viruses.102 103 104 2. Materials and methods 105 106 2.1. Mosquitoes 107 6To test the ability of our assays to detect vir...

Microfluidic systems for infectious disease diagnostics

“…490 Moutallier et al used a commercial microfluidic digital PCR system (Biomark™ HD, Standard BioTools, USA) for high-throughput detection of 64 mosquito-borne viruses in pools of mosquitoes from ZIKV endemic/epidemic areas. 505…”