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. 2020 Sep 14;20:282. doi: 10.1186/s12866-020-01966-6

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© The Author(s) 2020

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 3 — Sensitivity of CRISPR-Cas12a based assay in the detection of ASFV. a A qPCR positive blood sample (GD/GZ/0311) was subjected to serial (log10) dilution and examined by both the CRISPR-Cas12a and the commercial qPCR assay. Columns – CRISPR-Cas12a based assay; black dot – commercial qPCR assay. The dashed line represents the detection threshold for the virus. b A total of 17 qPCR positive blood were used for comparison using all three nucleic acid detection methods. Heatmap results represent the signal intensity of individual samples. c ASFV B646L Gene Plasmid Reference Material (5.8 × 10³ copies/μl) was subjected to serial dilution and examined by the CRISPR-Cas12a and the commercial qPCR. Columns – CRISPR-Cas12a based assay; black dot – commercial qPCR assay. Dashed line represents the detection threshold for the virus. d The panel shows that serial dilution of the positive sample (GD/GZ/0311) remained detectable over a 1 h time course with S/N ratio (fluorescence) measurements every 5 min. e The p72 genes of various ASFV genotypes I, II, VIII, IX, and X were synthesized and used to assess whether the CRISPR-Cas12a assay has the ability to test different ASFV genotypes. The dashed line represents the detection threshold for the virus. n = 3 technical replicates, bars represent mean ± SD