CRISPR-based rapid and ultra-sensitive diagnostic test for Mycobacterium tuberculosis

Rapid and simple-to-use diagnostic methods for tuberculosis are urgently needed. Recent development has unveiled the diagnostic power of the CRISPR system in the detection of viral infections. However, its potential use in detecting the Mycobacterium tuberculosis complex (MTB) remained unexplored. We developed a rapid CRISPR-based assay for TB detection and conducted a retrospective cohort study of 179 patients to evaluate the CRISPR-MTB test for identifying MTB in various forms of direct clinical samples. Its diagnostic performance was compared, in parallel with culture and the GeneXpert MTB/RIF assay (Xpert). The CRISPR-MTB test is highly sensitive with a near single-copy sensitivity, demands less sample input and offers shorter turnaround time than Xpert. When evaluated in the clinical cohort of both pulmonary and extra-pulmonary tuberculosis, the CRISPR-MTB test exhibited an overall improved sensitivity over both culture (79% vs 33%) and Xpert (79% vs 66%), without comprise in specificity (62/63, 98%). The CRISPR-MTB test exhibits an improved overall diagnostic performance over culture and Xpert across a variety of sample types, and offers great potential as a new diagnostic technique for both pulmonary and extra-pulmonary tuberculosis.

临床亟需快速易用的结核病诊断方法。近年来的研究进展揭示了CRISPR系统（CRISPR）在病毒感染检测中的诊断潜力。然而，其在检测结核分枝杆菌复合群（MTB）中的应用潜力尚未被探索。本研究开发了一种基于CRISPR的快速结核病检测方法，并纳入179名患者开展回顾性队列研究，以评估CRISPR-MTB检测法在多种直接临床样本中鉴定MTB的效果。该检测的诊断性能与培养法及GeneXpert MTB/RIF检测法（Xpert）进行了平行对比。CRISPR-MTB检测法具有极高的灵敏度，可达近乎单拷贝检测水平，且相较于Xpert，其所需样本投入量更少、检测周转时间更短。在纳入肺结核与肺外结核患者的临床队列中进行评估时，CRISPR-MTB检测法的整体灵敏度优于培养法（79% vs 33%）与Xpert（79% vs 66%），且特异性未受影响（62/63，98%）。在多种样本类型中，CRISPR-MTB检测法的整体诊断性能均优于培养法与Xpert，有望成为一种适用于肺结核与肺外结核的新型诊断技术。