Deployable Pan-Flavivirus and Pan-alphavirus Assays for Screening Pools of Medically Relevant Arthropod

摘要

CustomArray produces a field-deployable electrochemical microarray platform that is ideally suited to the multiplexed detection of pathogenic organisms. In our phase I effort we prototyped a complete detection process which includes: Probe design, sample prep, multiplexed PCR amplification, hybridization, detection and data analysis. We used our bioinformatic system called Genotyper to create an updated pan-flavivirus and pan-alphavirus detection system. We prototyped a detection system that relies on purification of viral nucleic acids directly from arthropods, primer directed amplification of these nucleic acids, hybridization of this material to microarray bound probes, detection of hybridization patterns, and finishes with an analysis that is able to accurately type the amplified material. Our pilot studies demonstrated the detection of 5 viral species, 2 Flaviviruses and 3 alphaviruses, with near single copy detection in a single reaction/detection assay. The detection device itself is hand-held and requires only a USB connection as its sole power source. Because we can use up to 2240 probes per assay, we are able to offer a resolution and redundancy that is unprecedented in a compact, field-deployable device, and will be able to expand our assay to all medically relevant arthropod-borne viruses.