1. A paper-based assay for detecting hypervirulent Klebsiella pnuemoniae using CRISPR-Cas13a system.
- Author
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Bhattacharjee, Gargi, Gohil, Nisarg, Khambhati, Khushal, Gajjar, Devarshi, Abusharha, Ali, and Singh, Vijai
- Subjects
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MEDICAL personnel , *KLEBSIELLA pneumoniae , *KLEBSIELLA , *DRUG resistance in microorganisms , *POLYSACCHARIDES , *CRISPRS - Abstract
[Display omitted] • A simple and user-friendly diagnostics to detect the hypervirulent K. pneumoniae. • CRISPR-assisted point-of-need tool that stands worthy of the ASSURED criteria. • Species-specific detection of solely the test-strains of K. pneumoniae. • No cross-reactivity observed when tested on different genera besides the test strains. • Instrument-free readout to extend the reach of the technology to remote locations. Klebsiella pneumoniae , a prevalent healthcare-associated pathogen, poses a significant challenge to diagnosis and treatment due to its virulence and antimicrobial resistance. The demand for more efficient, precise and accessible diagnostic methods is imperative, as current approaches are labor-intensive and resource-dependent. In this study, a CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-based diagnostic tool for rapid detection of hypervirulent K. pneumoniae infections was proposed. We integrated recombinase polymerase amplification (RPA) coupled with a lateral flow assay and Cas13a (CRISPR associated protein 13a) to target the housekeeping rpoB gene for species-specific detection and the capsular polysaccharide regulating gene rmpA for identification of hypervirulent strains of K. pneumoniae. Tested on 18 K. pneumoniae strains, the devised tool successfully detected hypervirulent strains K. pneumoniae M59 and K. pneumoniae KP109 showing presence of rmpA. This study allows to develop an instrument-free platform for routine diagnosis of K. pneumoniae from serum, urine, and saliva samples that would empower healthcare personnel to facilitate proper and timely treatment of infections caused by the K. pneumoniae. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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