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

• Published in: Nature communications Vol. 12; no. 1; p. 6018
• Main Authors: Kumar, Shailabh, Hol, Felix J. H., Pujhari, Sujit, Ellington, Clayton, Narayanan, Haripriya Vaidehi, Li, Hongquan, Rasgon, Jason L., Prakash, Manu
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 14.10.2021; Nature Publishing Group; Nature Portfolio
• Subjects: 631/1647/2163; 631/1647/277; 704/158/1469; Aedes; Animals; Biting; Culicidae; Deoxyribonucleic acid; Disease transmission; DNA; DNA viruses; Droplets; Female; High-Throughput Screening Assays; Humanities and Social Sciences; Insect bites; Insect Bites and Stings; Interrogation; Life Sciences; Low cost; Membranes; Microfluidics; Mosquito Vectors; Mosquitoes; multidisciplinary; Pathogens; Polymerase chain reaction; Ribonucleic acid; RNA; RNA viruses; Saliva; Science; Science (multidisciplinary); Vector-borne diseases; Viruses; Zika Virus; Zika Virus Infection
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-021-26300-0

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.