In:
PLOS Neglected Tropical Diseases, Public Library of Science (PLoS), Vol. 16, No. 8 ( 2022-8-29), p. e0010659-
Abstract:
Detection of Burkholderia pseudomallei , a causative bacterium for melioidosis, remains a challenging undertaking due to long assay time, laboratory requirements, and the lack of specificity and sensitivity of many current assays. In this study, we are presenting a novel method that circumvents those issues by utilizing CRISPR-Cas12a coupled with isothermal amplification to identify B . pseudomallei DNA from clinical isolates. Through in silico search for conserved CRISPR-Cas12a target sites, we engineered the CRISPR-Cas12a to contain a highly specific spacer to B . pseudomallei , named crBP34. The crBP34-based detection assay can detect as few as 40 copies of B . pseudomallei genomic DNA while discriminating against other tested common pathogens. When coupled with a lateral flow dipstick, the assay readout can be simply performed without the loss of sensitivity and does not require expensive equipment. This crBP34-based detection assay provides high sensitivity, specificity and simple detection method for B . pseudomallei DNA. Direct use of this assay on clinical samples may require further optimization as these samples are complexed with high level of human DNA.
Type of Medium:
Online Resource
ISSN:
1935-2735
DOI:
10.1371/journal.pntd.0010659
DOI:
10.1371/journal.pntd.0010659.g001
DOI:
10.1371/journal.pntd.0010659.g002
DOI:
10.1371/journal.pntd.0010659.g003
DOI:
10.1371/journal.pntd.0010659.g004
DOI:
10.1371/journal.pntd.0010659.s001
DOI:
10.1371/journal.pntd.0010659.s002
DOI:
10.1371/journal.pntd.0010659.s003
DOI:
10.1371/journal.pntd.0010659.s004
DOI:
10.1371/journal.pntd.0010659.s005
DOI:
10.1371/journal.pntd.0010659.s006
DOI:
10.1371/journal.pntd.0010659.s007
DOI:
10.1371/journal.pntd.0010659.s008
DOI:
10.1371/journal.pntd.0010659.s009
DOI:
10.1371/journal.pntd.0010659.s010
DOI:
10.1371/journal.pntd.0010659.s011
DOI:
10.1371/journal.pntd.0010659.r001
DOI:
10.1371/journal.pntd.0010659.r002
DOI:
10.1371/journal.pntd.0010659.r003
DOI:
10.1371/journal.pntd.0010659.r004
Language:
English
Publisher:
Public Library of Science (PLoS)
Publication Date:
2022
detail.hit.zdb_id:
2429704-5
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