Sensitive detection of a bacterial pathogen using allosteric probe-initiated catalysis and CRISPR-Cas13a amplification reaction

Jinjin Shen, Xiaoming Zhou, Yuanyue Shan, Huahua Yue, Ru Huang, Jiaming Hu () and Da Xing ()
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Jinjin Shen: MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, South China Normal University
Xiaoming Zhou: School of Life Sciences, South China Normal University
Yuanyue Shan: MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, South China Normal University
Huahua Yue: MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, South China Normal University
Ru Huang: MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, South China Normal University
Jiaming Hu: MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, South China Normal University
Da Xing: MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, South China Normal University

Nature Communications, 2020, vol. 11, issue 1, 1-10

Abstract: Abstract The ability to detect low numbers of microbial cells in food and clinical samples is highly valuable but remains a challenge. Here we present a detection system (called ‘APC-Cas’) that can detect very low numbers of a bacterial pathogen without isolation, using a three-stage amplification to generate powerful fluorescence signals. APC-Cas involves a combination of nucleic acid-based allosteric probes and CRISPR-Cas13a components. It can selectively and sensitively quantify Salmonella Enteritidis cells (from 1 to 105 CFU) in various types of samples such as milk, showing similar or higher sensitivity and accuracy compared with conventional real-time PCR. Furthermore, APC-Cas can identify low numbers of S. Enteritidis cells in mouse serum, distinguishing mice with early- and late-stage infection from uninfected mice. Our method may have potential clinical applications for early diagnosis of pathogens.

Date: 2020
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DOI: 10.1038/s41467-019-14135-9

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