An extraction-free one-pot assay for rapid detection of Klebsiella pneumoniae by combining RPA and CRISPR/Cas12a.
Biosens Bioelectron
; 267: 116740, 2024 Sep 05.
Article
in En
| MEDLINE
| ID: mdl-39244837
ABSTRACT
Klebsiella pneumoniae poses a significant threat to global public health. Traditional clinical diagnostic methods, such as bacterial culture and microscopic identification, are not suitable for point-of-care testing. In response, based on the suboptimal protospacer adjacent motifs, this study develops an extraction-free one-pot assay, named EXORCA (EXtraction-free One-pot RPA-CRISPR/Cas12a assay), designed for the immediate, sensitive and efficient detection of K. pneumoniae. The EXORCA assay can be completed within approximately 30 min at a constant temperature and allows for the visualization of results either through a fluorescence reader or directly by the naked eye under blue light. The feasibility of the assay was evaluated using twenty unextracted clinical samples, achieving a 100% (5/5) positive predictive value and a 100% (15/15) negative predictive value in comparison to qPCR. These results suggest that the EXORCA assay holds significant potential as a point-of-care testing tool for the rapid identification of pathogens, such as K. pneumonia.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Language:
En
Journal:
Biosens Bioelectron
Journal subject:
BIOTECNOLOGIA
Year:
2024
Document type:
Article
Affiliation country:
China
Country of publication:
Reino Unido