PT  - JOURNAL ARTICLE
AU  - Wu, Qianxin
AU  - Suo, Chenqu
AU  - Brown, Tom
AU  - Wang, Tengyao
AU  - Teichmann, Sarah A.
AU  - Bassett, Andrew R.
TI  - INSIGHT: a population scale COVID-19 testing strategy combining point-of-care diagnosis with centralised high-throughput sequencing
AID  - 10.1101/2020.06.01.127019
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.06.01.127019
4099  - http://biorxiv.org/content/early/2020/08/16/2020.06.01.127019.short
4100  - http://biorxiv.org/content/early/2020/08/16/2020.06.01.127019.full
AB  - We present INSIGHT (Isothermal NASBA-Sequencing-based hIGH-througput Test): a two-stage COVID-19 testing strategy, using a barcoded isothermal NASBA reaction that combines point-of-care diagnosis with next generation sequencing, aiming to achieve population-scale COVID-19 testing. INSIGHT combines the advantages of near-patient with centralised testing. Stage 1 allows a quick decentralised readout for early isolation of pre-symptomatic or asymptomatic patients. The same reaction products can then be used in a highly multiplexed sequencing-based assay in Stage 2, confirming the near-patient testing results and facilitating centralised data collection. Based on experiments using commercially acquired human saliva with spiked-in viral RNA as input, the INSIGHT platform gives Stage 1 results within one to two hours, using either fluorescence detection or a lateral flow (dipstick) readout, whilst simultaneously incorporating sample-specific barcodes into the amplification product. INSIGHT Stage 2 can be performed by directly pooling and sequencing all post-amplification barcoded Stage 1 products from hundreds of thousands of samples with minimal sample preparation steps. The 95% limit of detection (LoD-95) for INSIGHT is estimated to be below 50 copies of viral RNA per 20 μl of reaction. Our two-stage testing strategy is suitable for further development into a rapid home-based and point-of-care assay, and is potentially scalable to the population level.Competing Interest StatementQ.W., C.S., S.A.T. and A.R.B. are inventors on a patent application (GB2007724.4) submitted by Genome Research Limited, with the aim of making this technology freely available for research and deployment. In the past three years, S.A.T. has consulted for Genentech and Roche, and sits on Scientific Advisory Boards for Foresite Labs, Biogen and GlaxoSmithKline. T.B. is a consultant at ATDBio.