1. A microfluidic platform for highly parallel bite by bite profiling of mosquito-borne pathogen transmission
- Author
-
Felix J. H. Hol, Shailabh Kumar, Manu Prakash, Hongquan Li, Jason L. Rasgon, Sujit Pujhari, Clayton Ellington, Haripriya Vaidehi Narayanan, Stanford University, Interactions Virus-Insectes - Insect-Virus Interactions (IVI), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Centre de Recherche Interdisciplinaire / Center for Research and Interdisciplinarity [Paris, France] (CRI), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Pennsylvania State University (Penn State), Penn State System, This project was supported by grants to MP including NIH DP2-AI124336 New Innovator Award and USAID Grand Challenges: Zika and Future Threats Award. Microfluidic chip fabrications were done at Stanford Nano Shared Facilities (SNSF), supported by the National Science Foundation under award ECCS-1542152, at Stanford University and Institut Pasteur’s Biomaterials and Microfluidics core facility. F.J.H.H. was supported by a Rubicon fellowship for the Netherlands Foundation for Scientific Research, a Career Award at the Scientific Interface from the Burroughs Wellcome Fund, and a Marie Curie Fellowship from the European Union. S.P. and J.L.R. were supported by NIH Grants R01AI128201, R01AI150251, R01AI116636, USDA Hatch funds (Accession #1010032, Project #PEN04608), and a grant with the Pennsylvania Department of Health using Tobacco Settlement Funds. M.P. is supported by HHMI-Gates faculty scholarship and Chan Zuckerberg Biohub Investigator., European Project: 841893,H2020-MSCA-IF-2018,PiQiMosqBite(2019), Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Paris (UP), Hol, Felix, and Understanding the roles of pathogen infection and sensory cue integration in mosquito blood-feeding beh - PiQiMosqBite - - H2020-MSCA-IF-20182019-10-01 - 2021-11-07 - 841893 - VALID
- Subjects
[SDV]Life Sciences [q-bio] ,Microfluidics ,General Physics and Astronomy ,Membrane thickness ,Zika virus ,MESH: Microfluidics ,0302 clinical medicine ,Aedes ,MESH: Animals ,Pathogen ,0303 health sciences ,Multidisciplinary ,biology ,Lab-on-a-chip ,Transmission (medicine) ,Zika Virus Infection ,High-throughput screening ,MESH: Aedes ,3. Good health ,[SDV] Life Sciences [q-bio] ,MESH: Mosquito Vectors ,Female ,Sample collection ,MESH: High-Throughput Screening Assays ,Science ,030231 tropical medicine ,MESH: Zika Virus ,Mosquito Vectors ,General Biochemistry, Genetics and Molecular Biology ,Article ,03 medical and health sciences ,MESH: Zika Virus Infection ,MESH: Insect Bites and Stings ,parasitic diseases ,Animals ,MESH: Saliva ,Saliva ,030304 developmental biology ,Ecological epidemiology ,Insect Bites and Stings ,General Chemistry ,Zika Virus ,biology.organism_classification ,Virology ,High-Throughput Screening Assays ,Biting ,Culicidae ,MESH: Culicidae ,MESH: Female - Abstract
Mosquito bites transmit a number of pathogens via salivary droplets deposited during blood-feeding, resulting in potentially fatal diseases. Little is known about the genomic content of these nanodroplets, including the transmission dynamics of live pathogens. Here we introduce Vectorchip, a low-cost, scalable microfluidic platform enabling high-throughput molecular interrogation of individual mosquito bites. We introduce an ultra-thin PDMS membrane which acts as a biting interface to arrays of micro-wells. Freely-behaving mosquitoes deposit saliva droplets by biting into these micro-wells. By modulating membrane thickness, we observe species-dependent differences in mosquito biting capacity, utilizable for selective sample collection. We demonstrate RT-PCR and focus-forming assays on-chip to detect mosquito DNA, Zika virus RNA, as well as quantify infectious Mayaro virus particles transmitted from single mosquito bites. The Vectorchip presents a promising approach for single-bite-resolution laboratory and field characterization of vector-pathogen communities, and could serve as a powerful early warning sentinel for mosquito-borne diseases., High-throughput molecular surveillance of mosquitoes carrying dangerous pathogens is currently challenging. Here the authors present Vectorchip, a low-cost microfluidic platform enabling multiplexed detection of mosquito DNA, viral RNA and infectious viral particles at single bite resolution.
- Published
- 2021
- Full Text
- View/download PDF