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A streamlined CRISPR-based test for tuberculosis detection directly from sputum

SCIENCE ADVANCES. 2025-08; 
Alexandra G Bell, Owen R S Dunkley, Nisha H Modi, Yujia Huang, Soleil Tseng, Robert Reiss, Naranjargal Daivaa, J Lucian Davis, Deninson Alejandro Vargas, Padmapriya Banada, Yingda L Xie, Cameron Myhrvold Department of Molecular Biology, Princeton University, Princeton NJ 08544, USA.
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摘要

Mycobacterium tuberculosis (Mtb) is a major threat to global health, and there is an urgent need for affordable, simple tuberculosis (TB) diagnosis in underresourced areas. Here, we combine recombinase polymerase amplification with Cas13a and Cas12a detection to create two parallelized one-pot assays that detect two conserved elements of Mtb (IS6110 and IS1081) and a human DNA internal control. These assays are compatible with lateral flow and can be readily lyophilized. Our final assay showed a limit of detection of 69.0 CFU per milliliter for Mtb H37Rv and 80.5 CFU per milliliter for Mycobacterium bovis BCG in spiked sputum, with no cross-reactivity to diverse bacterial or fungal isolates. Clinical tests on 1... More

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