Mosquito bites transmit a number of human pathogens resulting in potentially fatal diseases including malaria, dengue, chikungunya, West Nile encephalitis, and Zika. Although female mosquitoes transmit pathogens via salivary droplets deposited during blood feeding on a host, very little is known about the genomic content of these nanoliter scale droplets, including the transmission dynamics of live pathogens. Here we introduce Vectorchip, a low-cost, scalable microfluidic platform for molecular interrogation of individual mosquito bites in a high-throughput fashion. An ultra-thin PDMS membrane coupled to a microfluidic chip acts as a biting interface, through which freely-behaving mosquitoes deposit saliva droplets by biting into isolated arrayed micro-wells enabling molecular interrogation of individual bites. By modulating membrane thickness, the device enables on-chip comparison of biting capacity and provides a mechanical filter allowing selection of a specific mosquito species. Utilizing Vectorchip, we show on-chip simultaneous detection of mosquito DNA as well as viral RNA from Zika infected Aedes aegypti mosquitoes - demonstrating multiplexed high-throughput screening of vectors and pathogens. Focus-forming assays performed on-chip quantify number of infectious viral particles transmitted during mosquito bites, enabling assessment of active virus transmission. The platform presents a promising approach for single-bite-resolution laboratory and field characterization of vector pathogen communities, to reveal the intricate dynamics of pathogen transmission, and could serve as powerful early warning artificial "sentinel" for mosquito-borne diseases.

中文翻译：

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Vectorchip：通过对蚊子传播的病原体进行咬合分析，实现高度平行咬合的微流控平台

蚊咬传播许多人类病原体，导致潜在的致命疾病，包括疟疾，登革热，基孔肯雅热，西尼罗河脑炎和寨卡病毒。尽管雌性蚊子通过在宿主供血期间沉积的唾液飞沫传播病原体，但对这些纳升级飞沫的基因组含量（包括活病原体的传播动力学）了解甚少。在这里，我们介绍Vectorchip，这是一种低成本，可扩展的微流体平台，用于以高通量方式对单个蚊虫叮咬进行分子询问。耦合到微流控芯片的超薄PDMS膜充当咬合界面，行为自由的蚊子通过咬入隔离的阵列微孔中而沉积唾液小滴，从而对单个咬合进行分子询问。通过调节膜厚度，该器件可以在芯片上比较咬合能力，并提供机械过滤器，可以选择特定的蚊子种类。利用Vectorchip，我们展示了片上同时检测Zika感染埃及伊蚊的蚊子DNA和病毒RNA的方法-展示了载体和病原体的多重高通量筛选。在芯片上执行的聚焦形成测定可量化蚊虫叮咬期间传播的传染性病毒颗粒的数量，从而能够评估主动病毒的传播。该平台为单病原实验室和媒介病原体群落的现场鉴定提供了一种有前途的方法，以揭示病原体传播的复杂动态，并可以作为蚊媒疾病的强大预警人工“前哨”。