Despite antibiotic treatments, tuberculosis (TB) remains an enormous global health burden, in part driven by the difficulties in timely and accurate diagnosis. Can we use CRISPR to improve TB diagnosis to make it faster, simpler, and more accessible? (1/21)

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)

Thank you to all the co-authors: Andrew Guo, Gordon Adams, Jacob E. Lemieux and Cameron Myhrvold @cambearon.bsky.social . We are also grateful to our funding sources, NIH and CDC.
Thank you for taking the time to follow along. Enjoy the paper📑! (11/11)

In summary, we developed SHINE-H5, SHINE-H5-CS, and SHINE-H7-Eurasian, addressing unmet needs in avian influenza surveillance, providing rapid, sensitive, specific detection suited for low-resource settings, with potential to strengthen outbreak response and preparedness☣️. (10/11)

We also designed and validated a Eurasian H7-specific SHINE assay (SHINE-H7-Eurasian), which reliably discriminated H7 Eurasian strains against North American H7 strains and unrelated seasonal influenza viruses. (9/11)

SHINE-H5-CS reliably discriminates between 2.3.4.4b and non-2.3.4.4b H5 sequences across a wide range of input concentrations. (8/11)

We developed a clade-specific SHINE assay （SHINE-H5-CS）specifically targeting clade 2.3.4.4b A(H5N1), which has been dominant in global H5 phylogeny since 2021. (7/11)

SHINE-AVIAN has a limit of detection of 121.7 copies/μL for H5N1 seedstocks through fluorescent readout and can detect as low as 25 copies/μL of H5N1 seedstocks through a lateral-flow readout. (6/11)

We developed SHINE-H5 for the detection of H5 avian influenza. SHINE-H5 demonstrated high specificity, showing strong signals only for H5 and none of non-target viruses or clinical flu-positive samples. (5/11)

In SHINE workflow, avian influenza genomes are reverse transcribed and amplified by SSIV, RPA, and T7. Complementary base pairing between Cas13-crRNA complex and amplified genome can activate Cas13, leading to a fluorescent or colorimetric signal through reporter cleavage. (4/11)

To improve field deployability, we used the SHINE (Streamlined Highlighting of Infections to Navigate Epidemics) platform, a one-pot CRISPR diagnostic technology optimized for point-of-need use. (3/11)
www.nature.com/articles/s41...

Current methods for avian flu diagnosis, including virus isolation, immunoassays, RT-PCR, and sequencing, either require substantial laboratory infrastructure or suffer from reduced sensitivity and specificity. (2/11)

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...