ArticleInfectious diseases of poverty2024
A novel single-tube LAMP-CRISPR/Cas12b method for rapid and visual detection of zoonotic Toxoplasma gondii in the environment.
Article in Infectious diseases of poverty, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed.
- Research Progress in Applications of the CRISPR-Cas12b System in Pathogen Nucleic Acid Detection.Current issues in molecular biology · 2026Review
- Critical appraisal of methodological limitations in adrenal tuberculosis outcome studies.International urology and nephrology · 2026Article
- Medium- and Large-sized Mammals on the Plateau: An Evolutionary Crucible for Zoonotic Pathogens - Ecological Drivers, Adaptive Evolution, and One Health Control.Microbial ecology · 2026Review
- LAMP-coupled CRISPR-Cas12a assays: A promising new tool for molecular diagnosis of leishmaniasis.PLoS neglected tropical diseases · 2026Article
- Next-generation molecular tools in veterinary parasitology: advances, challenges, and perspectives in the diagnosis of emerging parasites.Revista brasileira de parasitologia veterinaria = Brazilian journal of veterinary parasitology : Orgao Oficial do Colegio Brasileiro de Parasitologia Veterinaria · 2026Review
- Rapid and ultrasensitive detection ofFrontiers in cellular and infection microbiology · 2026Article
- Molecular diagnostics for cutaneous leishmaniasis: progress towards fulfilling the WHO target product profile.Parasitology · 2025Review
- Development of PCR, qPCR and LAMP methods for the detection ofFood and waterborne parasitology · 2025Article
- CRISPR-Cas-Based Diagnostics in Biomedicine: Principles, Applications, and Future Trajectories.Biosensors · 2025Review
- Leveraging CRISPR-Cas-Enhanced Isothermal Amplification Tools for Quick Identification of Pathogens Causing Livestock Diseases.Current microbiology · 2025Review
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Abstract
backgroundToxoplasma gondii oocysts, excreted in cat feces, pose a significant health risk to humans through contaminated soil and water. Rapid and accurate detection of T. gondii in environmental samples is essential for public health protection.
methodsWe developed a novel, single-tube detection method that integrates loop-mediated isothermal amplification (LAMP), the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12b system, and lateral flow immunoassay strips for rapid, visual identification of T. gondii. This method targets the T. gondii B1 gene, initially amplifies it with LAMP, directed by a single-guide RNA (sgRNA). It then recognizes the amplified target gene and activates trans-cleavage, cutting nearby single-stranded DNA (ssDNA) reporters. Fluorescence detection was performed using a 6-Carboxyfluorescein (FAM)-12N-Black Hole Quencher-1 (BHQ1) reporter, while Fluorescein Isothiocyanate (FITC)-12N-Biotin enabled visual detection on lateral flow strips. The method was tested for its ability to detect various T. gondii genotypes and related parasites, assessing its specificity and broad-spectrum applicability. It was further applied to real-world environmental samples to evaluate its practicality.
resultsThe LAMP-CRISPR/Cas12b method exhibited high specificity and broad-spectrum detection capability, successfully identifying nine T. gondii genotypes and distinguishing them from 11 other parasitic species. Sensitivity testing at both molecular (plasmid) and practical (oocyst) levels showed detection limits of 10 copies/μL and 0.1 oocyst, respectively. When applied to 112 environmental samples (soil, water, and cat feces), the method demonstrated 100% sensitivity, accurately reflecting known infection rates.
conclusionsThis LAMP-CRISPR/Cas12b single-tube method offers a robust, innovative approach for monitoring zoonotic T. gondii in environmental samples, with significant implications for public health surveillance.
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