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. 2019 Sep 27;11(10):904.
doi: 10.3390/v11100904.

A New High-Throughput Tool to Screen Mosquito-Borne Viruses in Zika Virus Endemic/Epidemic Areas

Sara Moutailler  1 , Lena Yousfi  2 , Laurence Mousson  3 , Elodie Devillers  4 , Marie Vazeille  5 , Anubis Vega-Rúa  6 , Yvon Perrin  7 , Frédéric Jourdain  8   9 , Fabrice Chandre  10   11 , Arnaud Cannet  12 , Sandrine Chantilly  13 , Johana Restrepo  14 , Amandine Guidez  15 , Isabelle Dusfour  16 , Filipe Vieira Santos de Abreu  17 , Taissa Pereira Dos Santos  18 , Davy Jiolle  19 , Tessa M Visser  20 , Constantianus J M Koenraadt  21 , Merril Wongsokarijo  22 , Mawlouth Diallo  23 , Diawo Diallo  24 , Alioune Gaye  25 , Sébastien Boyer  26 , Veasna Duong  27 , Géraldine Piorkowski  28 , Christophe Paupy  29 , Ricardo Lourenco de Oliveira  30 , Xavier de Lamballerie  31 , Anna-Bella Failloux  32
Affiliations

A New High-Throughput Tool to Screen Mosquito-Borne Viruses in Zika Virus Endemic/Epidemic Areas

Sara Moutailler et al. Viruses. .

Abstract

Mosquitoes are vectors of arboviruses affecting animal and human health. Arboviruses circulate primarily within an enzootic cycle and recurrent spillovers contribute to the emergence of human-adapted viruses able to initiate an urban cycle involving anthropophilic mosquitoes. The increasing volume of travel and trade offers multiple opportunities for arbovirus introduction in new regions. This scenario has been exemplified recently with the Zika pandemic. To incriminate a mosquito as vector of a pathogen, several criteria are required such as the detection of natural infections in mosquitoes. In this study, we used a high-throughput chip based on the BioMark™ Dynamic arrays system capable of detecting 64 arboviruses in a single experiment. A total of 17,958 mosquitoes collected in Zika-endemic/epidemic countries (Brazil, French Guiana, Guadeloupe, Suriname, Senegal, and Cambodia) were analyzed. Here we show that this new tool can detect endemic and epidemic viruses in different mosquito species in an epidemic context. Thus, this fast and low-cost method can be suggested as a novel epidemiological surveillance tool to identify circulating arboviruses.

Keywords: microfluidic analysis; molecular epidemiology; mosquito-borne viruses; surveillance.

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Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Figures

Figure 1
Figure 1
BioMark™ dynamic array system specificity test (96.96 chip). Specificity of primers/probe sets from the Table S1 are presented in two figures, (A,B). Fifty-one sets of primers/probe targeting viruses are presented in (A) and 94 sets in (B), some of them are present into both figures. Each square corresponds to a single real-time PCR reaction, where rows indicate the pathogen in the positive control and columns represent the targets of each primer/probe set. Ct values for each reaction are indicated in color; the corresponding color scale is presented in the legend on the right. The darkest shade of blue and black squares are considered as negative reactions with Ct > 30. chik: Chikungunya; deng: Dengue; EnM: Murray Encephalitis; St Louis Enc: Saint Louis Encephalitis; EEE: Eastern Equine Encephalitis; WEE: Western Equine Encephalitis; VEE: Venezuelan Equine Encephalitis; JE: Japanese Encephalitis; YF: Yellow fever; Kok: Kokobera group; Maya: Mayaro; ONN: O’nyong nyong; RVF: Rift Valley Fever; Seml: Semliki forest; Spond: Spondweni; Tahy: Tahyna; WN: West Nile; Wess: Wesselsbron; Kout: Koutango; Guama: Guama Group; Calif Enc: California Encephalitis; GpC: C Group; Key: Keystone; Snow: Snowshoe hare.
Figure 1
Figure 1
BioMark™ dynamic array system specificity test (96.96 chip). Specificity of primers/probe sets from the Table S1 are presented in two figures, (A,B). Fifty-one sets of primers/probe targeting viruses are presented in (A) and 94 sets in (B), some of them are present into both figures. Each square corresponds to a single real-time PCR reaction, where rows indicate the pathogen in the positive control and columns represent the targets of each primer/probe set. Ct values for each reaction are indicated in color; the corresponding color scale is presented in the legend on the right. The darkest shade of blue and black squares are considered as negative reactions with Ct > 30. chik: Chikungunya; deng: Dengue; EnM: Murray Encephalitis; St Louis Enc: Saint Louis Encephalitis; EEE: Eastern Equine Encephalitis; WEE: Western Equine Encephalitis; VEE: Venezuelan Equine Encephalitis; JE: Japanese Encephalitis; YF: Yellow fever; Kok: Kokobera group; Maya: Mayaro; ONN: O’nyong nyong; RVF: Rift Valley Fever; Seml: Semliki forest; Spond: Spondweni; Tahy: Tahyna; WN: West Nile; Wess: Wesselsbron; Kout: Koutango; Guama: Guama Group; Calif Enc: California Encephalitis; GpC: C Group; Key: Keystone; Snow: Snowshoe hare.
Figure 2
Figure 2
Screening of artificially infected mosquitoes through the BioMark™ dynamic array system developed (96.96 chip). Each square corresponds to a single real-time PCR reaction, where rows indicate the batches of mosquitoes tested and columns represent the targets of each primer/probe set. Crosses indicate cross-reaction of assays. Ct values for each reaction are indicated in color; the corresponding color scale is presented in the legend on the right. The darkest shade of blue and black squares are considered as negative reactions with Ct > 30. chik: Chikungunya; deng: Dengue; EnM: Murray Encephalitis; St Louis Enc: Saint Louis Encephalitis; EEE: Eastern Equine Encephalitis virus; WEE: Western Equine Encephalitis; VEE: Venezuelan Equine Encephalitis; JE: Japanese Encephalitis; YF: Yellow fever; Kok: Kokobera grou; Maya: Mayaro; ONN: O’nyong nyong; RVF: Rift Valley Fever; Seml: Semliki fores; Spond: Spondweni; Tahy: Tahyna; WN: West Nile; Wess: Wesselsbron; Kout: Koutango.
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