Journal: Poultry Science

Article Title: Field-deployable multiplex RAA-CRISPR/Cas12a platform rapidly and simultaneously detects seven Eimeria species in chickens

doi: 10.1016/j.psj.2026.106681

Figure Lengend Snippet: Screening of species-specific crRNAs and determination of fluorescence and visual detection thresholds for the E-MRC12a assay. (A-G) Real-time fluorescence curves of CRISPR/Cas12a reactions targeting the 18S rDNA of the seven Eimeria species using three candidate crRNAs per species: (A) E. maxima (EM1-EM3), (B) E. acervulina (EA1-EA3), (C) E. necatrix (EN1-EN3), (D) E. mitis (EI1-EI3), (E) E. praecox (EP1-EP3), (F) E.brunelti (EB1-EB3) and (G) E. tenella (ET1). (H) UV images of CRISPR/Cas12a reaction products for each species-specific crRNA. (I) Determination of the visual fluorescence threshold. NT: no-template control. Error bars represent mean ± SD (n = 3 per group).

Article Snippet: The CRISPR/Cas12a trans-cleavage assay was performed in a 20 μL reaction containing 2.5 μL of 10 × NEBuffer, 2 μL of LbCas12a protein (1 μM), 1 μL of crRNA (1 μM), 2 μL of ssDNA reporter (10 μM), 2 μL of RAA product, and ddH2O to volume.

Techniques: Fluorescence, CRISPR, Control

Journal: Poultry Science

Article Title: Field-deployable multiplex RAA-CRISPR/Cas12a platform rapidly and simultaneously detects seven Eimeria species in chickens

doi: 10.1016/j.psj.2026.106681

Figure Lengend Snippet: Evaluation of the specificity, sensitivity, and repeatability of the E-MRC12a assay . (A) Heatmap of fluorescence intensities from CRISPR/Cas12a reactions using species-specific crRNAs targeting genomic DNA from the seven Eimeria species, Escherichia coli, Salmonella spp ., and confirmed Eimeria -negative fecal samples. (B) Visual fluorescence under 254 nm UV light corresponding to (A). (C) Heatmap showing the sensitivity evaluation of the E-MRC12a assay. (D) Visual fluorescence under 254 nm UV light corresponding to (C). (E) Heatmap illustrating repeatability assessment of the E-MRC12a assay. NT: No-template control. For panels (A) and (C), different letters within a row indicate statistically significant differences ( P < 0.05); identical letters indicate no significant difference ( P > 0.05).

Article Snippet: The CRISPR/Cas12a trans-cleavage assay was performed in a 20 μL reaction containing 2.5 μL of 10 × NEBuffer, 2 μL of LbCas12a protein (1 μM), 1 μL of crRNA (1 μM), 2 μL of ssDNA reporter (10 μM), 2 μL of RAA product, and ddH2O to volume.

Techniques: Fluorescence, CRISPR, Control

Journal: Poultry Science

Article Title: Field-deployable multiplex RAA-CRISPR/Cas12a platform rapidly and simultaneously detects seven Eimeria species in chickens

doi: 10.1016/j.psj.2026.106681

Figure Lengend Snippet: Schematic view of the nucleic acid extraction and Eimeria –Multiplex RAA–CRISPR/Cas12a (E-MRC12a) typing assay.

Article Snippet: The CRISPR/Cas12a trans-cleavage assay was performed in a 20 μL reaction containing 2.5 μL of 10 × NEBuffer, 2 μL of LbCas12a protein (1 μM), 1 μL of crRNA (1 μM), 2 μL of ssDNA reporter (10 μM), 2 μL of RAA product, and ddH2O to volume.

Techniques: Extraction, Multiplex Assay, CRISPR