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. 2022 Nov 3;13:977382. doi: 10.3389/fmicb.2022.977382

Figure 2.

Figure 2

The ultrasensitive detection assay based on CRISPR/Cas13a. (A) The designed LwaCas13a-crRNAs were used to detect the targets of L gene that was amplified by primer mix. The Cas13-L-crRNA1 and Cas13-L-crRNA2 exhibited stronger signals using the same protocol. Values represent mean ± SD (n = 3). (B) The specificity of Cas13-L-crRNA1 and Cas13-L-crRNA2. Several hemorrhagic fever viral RNA was detected parallelly using Cas13-L-crRNA1 or Cas13-L-crRNA2. The experimental results showed that there was no cross-reactivity in the detection. Values represent mean ± SD (n = 3). (C) The sensitivity of Cas13a-based assay using Cas13-L-crRNA1. The diluted SFTSV RNA standards were used as detection targets. (n = 3). (D) The Venn diagram showed the concordance of the results of the three assay systems on clinical samples. (E) The fluorescence visualization of the Cas13a-based assay for detection of clinical samples. A total of 52 samples were used for the evaluation. The results were observed and took pictures at 20 min for the Cas13a reaction under ultraviolet light (n = 3). PC, positive control; NC, no-template control.