Detection of Clostridium perfringens Using Novel Methods Based on Recombinase-Aided Amplification Assay-Assisted CRISPR/Cas12a System

Figure 6

Evaluation of the practicability of RAA-CRISPR/Cas12a-based methods for detecting C. perfringens in clinical and spiked samples. Genomic DNAs were extracted from the tissues of two abnormal death Milu and then were detected using RAA-CRISPR/Cas12a-FL (a), RAA-CRISPR/Cas12a-LFS (b), and qPCR (c) assays. 1–6 were lung, heart, spleen, kidney, liver, and jejunum, respectively, collected from the Milu dying of C. perfringens; 7–12 were lung, heart, spleen, kidney, liver, and jejunum, respectively, collected from the Milu dying of non-C. perfringens pathogen. (d and e) Human blood and fecal samples were collected and used to evaluate the practicability of RAA-CRISPR/Cas12a-based methods in the diagnosis of patients infected with C. perfringens. Blood samples and fecal samples were contaminated with 1 × 10³ CFU C. perfringens, and then the spiked samples and normal samples were detected using RAA-CRISPR/Cas12a-FL (d) and RAA-CRISPR/Cas12a-LFS (e) methods. 1, spiked blood sample; 2, normal blood sample; 3, spiked fecal sample; 4, normal blood sample. n = 3 technical replicates; two-tailed Student’s t test; ; bars represent mean ± SEM.

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