A microfluidic platform for highly parallel bite by bite profiling of mosquito-borne pathogen transmission

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.

Barcoded Asaia bacteria enable mosquito in vivo screens and identify novel systemic insecticides and inhibitors of malaria transmission

“…Sampling intensity of Asaia encoded blood meals differed between wells, and some wells were sampled by more than 1 mosquito. This is in line with the biting behaviour observed with the Vectorchip that contains arrayed glucose meals to sample saliva from individual mosquito bites [37]. By using a surplus of mosquitoes relative to the number of blood meals, we ensured that every well was sampled by multiple mosquitoes.…”

Exploratory phosphoproteomics profiling of Aedes aegypti Malpighian tubules during blood meal processing reveals dramatic transition in function

“…This problem could be solved with the use of a blood feeding system allowing for blood feeding and sampling of individual mosquitoes. Kumar et al recently described a 3D-printed chip system that allows for feeding and monitoring of individual mosquitoes [79], which would prove useful for future experiments to assay for rapid changes in protein phosphorylation within the first hour of blood feeding.…”

Nucleic Acid Amplification‐Based Technologies (NAAT)—Toward Accessible, Autonomous, and Mobile Diagnostics

“…Fabrication of a silicone membrane integrated microfluidic chip c) Autonomous sample loading via mosquito bites and d) on‐chip PCR to detect mosquito‐borne pathogens. Reproduced under the terms of the CC BY 4.0 license [ 11 ] Copyright 2021, The Authors, published by Nature publishing group.…”

“…[10] On-chip saliva collection directly from freely-behaving mosquitoes and extraction-free PCR in multiple wells for detection of Zika virus in salivary droplets was demonstrated (Figure 3b-d). [11] The chip utilized a skin-mimic silicone membrane on a PDMS chip, and utilized attractors so that mosquitoes were drawn to the chip, and bit through the membrane depositing their saliva directly into the reaction wells. This work showed that miniaturized chips using NAAT can potentially be used in the field for vector-borne pathogen surveillance to prevent infections in human populations.…”

“…[ 6,7 ] Among these, NAATs provide a very high degree of accuracy and sensitivity enabling detection of pathogens at extremely low concentrations. [ 8 ] NAATs are also important for other targets including liquid biopsies for cancers, [ 9 ] sexually transmitted infections (STIs), [ 10 ] vector‐borne diseases, [ 11 ] drug resistance in pathogens, [ 12 ] environmental DNA for surveillance of invasive species, [ 13 ] and early detection of agricultural pests and pathogens. [ 14,15 ]…”