TY - JOUR
T1 - A microfluidic platform for highly parallel bite by bite profiling of mosquito-borne pathogen transmission
AU - Kumar, Shailabh
AU - Hol, Felix J.H.
AU - Pujhari, Sujit
AU - Ellington, Clayton
AU - Narayanan, Haripriya Vaidehi
AU - Li, Hongquan
AU - Rasgon, Jason L.
AU - Prakash, Manu
N1 - Funding Information:
This project was supported by grants to MP including NIH DP2-AI124336 New Innovator Award and USAID Grand Challenges: Zika and Future Threats Award. Micro-fluidic 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. We thank all members of Prakash Lab for useful feedback on the work. We thank Louis Lambrechts (Insect Virus Interactions Unit, Institut Pasteur, Paris, France) and Lark Coffey (School of Veterinary Medicine, University of California, Davis, CA, USA) for providing access to arbovirus laboratories.
Publisher Copyright:
© 2021, The Author(s).
PY - 2021/12/1
Y1 - 2021/12/1
N2 - 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.
AB - 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.
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U2 - 10.1038/s41467-021-26300-0
DO - 10.1038/s41467-021-26300-0
M3 - Article
C2 - 34650045
AN - SCOPUS:85117400421
SN - 2041-1723
VL - 12
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 6018
ER -