ArticlePLoS neglected tropical diseases2022
Highly specific and sensitive detection of Burkholderia pseudomallei genomic DNA by CRISPR-Cas12a.
Article in PLoS neglected tropical diseases, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed.
- An Fe single-atom nanozyme-sensitized RPA-CRISPR/Cas12a biosensor for the early detection of potential microcystins-producing cyanobacteria.Mikrochimica acta · 2026Article
- CRISPR-based environmental detection of Burkholderia pseudomallei identifies sanitation gaps and melioidosis risk in northeast Thailand.Nature communications · 2026Article
- Current Melioidosis Diagnostic Landscape and Missed Opportunities in Biomarker Development.Diagnostics (Basel, Switzerland) · 2026Review
- Construction of an RPA-CRISPR visualization system for rapid detection of Burkholderia mallei.BMC microbiology · 2026Article
- Laboratory diagnosis of melioidosis.PLoS neglected tropical diseases · 2025Review
- Development of a field-deployable RPA-CRISPR/Cas12a assay for the detection of Cyclospora cayetanensis in human feces.Parasites & vectors · 2025Article
- Whole genome study and construction of SHERLOCK detection method for endemic strains ofMicrobiology spectrum · 2025Article
- A novel system with robust compatibility and stability for detectingMicrobiology spectrum · 2024Article
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Authors and funding
12 authors.
Funding
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.
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Registered trials
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