Journal
ACS NANO
Volume 14, Issue 2, Pages 2497-2508Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acsnano.0c00022
Keywords
lateral flow assay; CRISPR/Cas9; Listeria monocytogenes detection; GMOs detection; ASFV detection
Categories
Funding
- National Natural Science Foundation of China [21475048, 21874049, 91959128, 21904042, 31941004]
- National Science Fund for Distinguished Young Scholars of Guangdong Province [2014A030306008]
- Key-Area Research and Development Program of Guangdong Province [2019B020211004]
- National Key Research and Development Program of China [2016YFD0501300]
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The lateral flow assay is one of the most convenient analytical techniques for analyzing the immune response, but its applicability to precise genetic analyses is limited by the false-positive signal and tedious and inefficient hybridization steps. Here, we introduce the CRISPR (clustered regularly interspaced short palindromic repeats) /Cas system into the lateral flow assay, termed CRISPR/Cas9-mediated lateral flow nucleic acid assay (CASLFA), to address such issues. In this study, CASLFA is utilized to identify Listeria monocytogenes, genetically modified organisms (GMOs), and African swine fever virus (ASFV) at a detection limit of hundreds of copies of genome samples with high specificity within 1 h. We further evaluated the performance of CASLFA in a nonlaboratory environment and successfully confirmed 27 ASFV-infected samples from 110 suspected swine serum samples, with an accuracy of 100% when compared to real-time PCR (RT-PCR) assay. CASLFA satisfies some of the characteristics of a next-generation molecular diagnostics tool due to its rapidity and accuracy, allowing for point-of-care use without the need for technical expertise and complex ancillary equipment. This method has great potential for gene analysis in resource-poor or nonlaboratory environments.
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