Abstract
ABSTRACT 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.
Highlights
Tuberculosis (TB) is the leading cause of death among infectious diseases, with higher mortality than HIV and malaria worldwide [1]
Highly sensitive and simple-touse Mycobacterium tuberculosis complex (MTB) assay by combining a Recombinase Polymerase Amplification (RPA) reaction with a Clustered regularly interspaced short palindromic repeat (CRISPR)/Cas12a step for target detection (Figure 1)
To establish a rapid and simple-to-use extraction method for the CRISPR-MTB assay, we set out to optimize the extraction process based on a combination strategy of beads beating, chemical lysis and heating to ensure high efficiency of MTB DNA extraction
Summary
Tuberculosis (TB) is the leading cause of death among infectious diseases, with higher mortality than HIV and malaria worldwide [1]. As rapid and accurate diagnosis of tuberculosis is a prerequisite for effective treatment, alternative methods which allow rapid screening and diagnostic are urgently needed
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
