ArticlePLoS neglected tropical diseases2022
Rapid and sensitive point-of-care detection of Leptospira by RPA-CRISPR/Cas12a targeting lipL32.
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 30 papers, 1 of them a synthesis that pooled it.
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Who cites it
30 citing papers in PubMed, 1 synthesis or guideline pooled it, 76 citations in OpenAlex.
- Trends and advances inFrontiers in microbiology · 2024Pooled it
- Diagnosis of leptospirosis in animals: challenges and perspectives.Journal of clinical microbiology · 2026Review
- Early diagnosis and treatment of leptospirosis: Optimizing clinical outcomes.The Journal of infection · 2026Review
- Development of a field-deployable RPA-CRISPR/Cas12a assay for the detection of Cyclospora cayetanensis in human feces.Parasites & vectors · 2025Article
- Nanoparticle-based biosensors coupled with loop-mediated isothermal amplification for rapid, visual, and in-field identification of pathogenic Leptospira.BMC infectious diseases · 2025Article
- CRISPR-Cas12 Application for the Detection ofInternational journal of molecular sciences · 2025Article
- Development of Reverse Transcriptase Recombinase Polymerase Amplification Combined with Lateral Flow Dipstick for Rapid Detection of Tilapia Lake Virus (TiLV): Pilot Study.Veterinary sciences · 2025Article
- Leveraging CRISPR-Cas-Enhanced Isothermal Amplification Tools for Quick Identification of Pathogens Causing Livestock Diseases.Current microbiology · 2025Review
- Conventional and cutting-edge advances in plant virus detection: emerging trends and techniques.3 Biotech · 2025Review
- Leptospirosis-Improving Healthcare Outcomes for a Neglected Tropical Disease.Open forum infectious diseases · 2025Review
- Integrated CRISPR-Cas12a and RAA one-pot visual strategy for the rapid identification ofFrontiers in cellular and infection microbiology · 2025Article
- Rapid detection of feline parvovirus using RAA-CRISPR/Cas12a-based lateral flow strip and fluorescence.Frontiers in microbiology · 2025Article
- Rapid detection of FAdV-4 by one-tube RPA-CRISPR/Cas12a assay.Frontiers in microbiology · 2025Article
- Point-of-care test of blood Plasmodium RNA within a Pasteur pipette using a novel isothermal amplification without nucleic acid purification.Infectious diseases of poverty · 2024Article
- Production and characterization of immunoglobulin G anti-rLipL32 antibody as a biomarker for the diagnosis of leptospirosis.Veterinary world · 2024Article
- A multiplex RPA-CRISPR/Cas12a-based POCT technique and its application in human papillomavirus (HPV) typing assay.Cellular & molecular biology letters · 2024Article
- Ov-RPA-CRISPR/Cas12a assay for the detection of Opisthorchis viverrini infection in field-collected human feces.Parasites & vectors · 2024Article
- A Method for Detecting Five Carbapenemases in Bacteria Based on CRISPR-Cas12a Multiple RPA Rapid Detection Technology.Infection and drug resistance · 2024Article
- RPA-CRISPR-Cas13a-assisted detection method of transmissible gastroenteritis virus.Frontiers in veterinary science · 2024Article
- Article
Corrections and comments
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Authors and funding
10 authors at 2 institutions in 3 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundOne of the key barriers preventing rapid diagnosis of leptospirosis is the lack of available sensitive point-of-care testing. This study aimed to develop and validate a clustered regularly-interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 12a (CRISPR/Cas12a) platform combined with isothermal amplification to detect leptospires from extracted patient DNA samples. METHODOLOGY/PRINCIPAL
findingsA Recombinase Polymerase Amplification (RPA)-CRISPR/Cas12a-fluorescence assay was designed to detect the lipL32 gene of pathogenic Leptospira spp. The assays demonstrated a limit of detection (LOD) of 100 cells/mL, with no cross-reactivity against several other acute febrile illnesses. The clinical performance of the assay was validated with DNA extracted from 110 clinical specimens and then compared to results from qPCR detection of Leptospira spp. The RPA-CRISPR/Cas12a assay showed 85.2% sensitivity, 100% specificity, and 92.7% accuracy. The sensitivity increased on days 4-6 after the fever onset and decreased after day 7. The specificity was consistent for several days after the onset of fever. The overall performance of the RPA-CRISPR/Cas12a platform was better than the commercial rapid diagnostic test (RDT). We also developed a lateral flow detection assay (LFDA) combined with RPA-CRISPR/Cas12a to make the test more accessible and easier to interpret. The combined LFDA showed a similar LOD of 100 cells/mL and could correctly distinguish between known positive and negative clinical samples in a pilot study. CONCLUSIONS/SIGNIFICANCE: The RPA-CRISPR/Cas12 targeting the lipL32 gene demonstrated acceptable sensitivity and excellent specificity for detection of leptospires. This assay might be an appropriate test for acute leptospirosis screening in limited-resource settings.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.