Avian flu threatens global health and agriculture, with pandemic potential☣️ if it spreads between humans😷. To enable surveillance, we developed 3 CRISPR-based assays for rapid subtype- and clade-specific detection of avian influenza in low-resource settings.⬇️ (1/11)
www.medrxiv.org/content/10.1...

Thank you to all the co-authors: Ryungeun Song, Yujia Huang, Soonwoo Hong, Ibrahim Motlani, Howard A. Stone and Cameron Myhrvold. We are also grateful to our funding sources, NIH, CDC and Princeton Catalysis Initiative (PCI).

Thank you for taking the time to follow along, enjoy the paper! (12/12)

mSHINE enables the rapid on-site surveillance of clinically-relevant mutations and pathogens with unprecedented ease, speed and breadth, allowing for distributed multiplexed testing in resource-limited settings that are inaccessible to existing technologies. (11/12)

Lung cancer is the leading global cause of cancer-related mortality and 85% of lung cancer cases are non-small cell lung cancer (NSCLC ). We demonstrated detection of the EGFR exon 19 deletions and the critical L858R SNP implicated in NSCLC drug response. (10/12)

Combining the comb and valve geometries, we developed a 112-plex configuration. We demonstrated its specificity by generating this striped pattern on chip. (9/12)

We showed that mSHINE is as sensitive as standard SHINE. (8/12)

We showed that mSHINE is specific, without cross-talk between chambers, by generating this staggered checkerboard pattern on chip. (7/12)

We incorporated droplet lyophilization and an on-chip mixing process into the device. (6/12)

We prototyped three flow distribution geometries, a tree, comb and valve geometry, each with a different principle to enable even flow distribution. The tree geometry relies on equal path length, the comb, on adjusted hydraulic resistances and the valve, on surface-tension-regulated valves. (5/12)

We develop mSHINE, using microfluidics to spatially multiplex SHINE to enable >100-plex nucleic acid detection. (4/12)

During the pandemic our lab developed SHINE (Streamlined Highlighting of Infections to Navigate Epidemics), a sensitive and deployable CRISPR-based technology for nucleic acid detection. But like many other deployable approaches, SHINE is difficult to multiplex. (3/12)

Existing CRISPR-based technologies are either highly multiplexed or simple/portable.

Simple (few-plex) methods
•SHERLOCK
•DETECTR
•HOLMES
•SHINE
•STOPCovid
•miSHERLOCK
•Many others…

Highly multiplexed methods
•CARMEN
•mCARMEN
•MiCar
•bbCARMEN

(2/12)

Multiplexed nucleic acid detection is critical for the surveillance of pathogens and mutations, but existing technologies either require extensive lab equipment or have low multiplexing. We built a portable microfluidic manifold for massively multiplexed CRISPR-based detection. (1/12)