Department of Bioengineering, Stanford University, Stanford, CA, USA.
Insect Virus Interactions Unit, Department of Virology, Institut Pasteur, UMR2000, CNRS, Paris, France.
Nat Commun. 2021 Oct 14;12(1):6018. 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.
蚊子叮咬通过吸血时留下的唾液滴传播多种病原体，导致潜在致命疾病。关于这些纳米液滴的基因组内容，包括活病原体的传播动态，人们知之甚少。在这里，我们介绍了 Vectorchip，这是一种低成本、可扩展的微流控平台，可实现对单个蚊子叮咬的高通量分子检测。我们引入了一种超薄 PDMS 膜，该膜充当 biting interface 以形成微井阵列。自由活动的蚊子通过咬入微井来分泌唾液滴。通过调节膜的厚度，我们观察到蚊子叮咬能力的种间差异，可用于选择性样本收集。我们展示了 RT-PCR 和焦点形成测定法，以检测蚊子 DNA、寨卡病毒 RNA，以及量化从单个蚊子叮咬传播的传染性马亚罗病毒颗粒。Vectorchip 为单咬分辨率的实验室和野外特征向量病原体群落提供了一种很有前途的方法，并可以作为蚊虫传播疾病的强大预警哨兵。
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