Isothermal RNA Amplification for the Detection of Viable Pathogenic Bacteria to Estimate the Salmonella Virulence for Causing Enteritis

Xue, Ting; Lü, Ying; Yang, Hao; Hu, Xinyue; Zhang, Kaixiang; Ren, Yao; Wu, Chengyong; Xia, Xuhan; Deng, Ruijie; Wang, Yuxi

doi:10.1021/acs.jafc.1c07182

Cited by 40 publications

(22 citation statements)

References 37 publications

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“…The method showed a satisfactory sensitivity compared with qPCR (LOD = 10 4 CFU/mL). In addition, an isothermal RNA amplification (NASBA) was coupled with the CRISPR‐Cas13a system, termed as cNASBA assay, which could detect as low as 1 CFU and 1% viable bacteria detectable within 2.5 h (Xue et al., 2022). Remarkably, compared to qPCR, the cNASBA assay can quantify viable S. enterica and thus can improve the accuracy of virulence estimation.…”

Section: Applications Of Crispr‐cas‐based Detection In Food Safety Mo...mentioning

confidence: 99%

CRISPR‐Cas‐based detection for food safety problems: Current status, challenges, and opportunities

Man

Liu

2022

Comp Rev Food Sci Food Safe

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Food safety is one of the biggest public issues occurring around the world. Microbiological, chemical, and physical hazards can lead to food safety issues, which may occur at all stages of the supply chain. In order to tackle food safety issues and safeguard consumer health, rapid, accurate, specific, and field-deployable detection methods meeting diverse requirements are one of the imperative measures for food safety assurance. CRISPR-Cas system, a newly emerging technology, has been successfully repurposed in biosensing and has demonstrated huge potential to establish conceptually novel detection methods with high sensitivity and specificity. This review focuses on CRISPR-Cas-based detection and its current status and huge potential specifically for food safety inspection. We firstly illustrate the pending problems in food safety and summarize the popular detection methods. We then describe the potential applications of CRISPR-Casbased detection in food safety inspection. Finally, the challenges and futuristic

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Section: Applications Of Crispr‐cas‐based Detection In Food Safety Mo...mentioning

confidence: 99%

CRISPR‐Cas‐based detection for food safety problems: Current status, challenges, and opportunities

Man

Liu

2022

Comp Rev Food Sci Food Safe

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“…The APC-Cas assay could detect as low as 1 CFU/ml of S. enteritidis and exhibited superior specificity because of the dual recognition and triple amplification processes. Moreover, detection of viable bacteria is essential for bacterial virulence; therefore, one study developed a CRISPR/Cas13-based NASBA assay (called the cNASBA assay) that couples NASBA with the Cas13a reaction to detect and quantify viable S. enterica within 2.5 h, with an LOD of 1.5 CFU/ml of viable bacteria ( Xue et al, 2022 ). In addition, Cas13a-based detection combined with PCR amplification was performed to detect S. aureus , another pathogen of foodborne diseases, exhibiting an LOD of 1 CFU/ml within 4 h ( Zhou J. et al, 2020 ).…”

Section: Cas13a-based Detection Of Bacteria and Other Pathogensmentioning

confidence: 99%

CRISPR-Cas13a system: A novel tool for molecular diagnostics

Zhao

Qiu

et al. 2022

Front. Microbiol.

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The clustered regularly interspaced short palindromic repeats (CRISPR) system is a natural adaptive immune system of prokaryotes. The CRISPR-Cas system is currently divided into two classes and six types: types I, III, and IV in class 1 systems and types II, V, and VI in class 2 systems. Among the CRISPR-Cas type VI systems, the CRISPR/Cas13a system has been the most widely characterized for its application in molecular diagnostics, gene therapy, gene editing, and RNA imaging. Moreover, because of the trans-cleavage activity of Cas13a and the high specificity of its CRISPR RNA, the CRISPR/Cas13a system has enormous potential in the field of molecular diagnostics. Herein, we summarize the applications of the CRISPR/Cas13a system in the detection of pathogens, including viruses, bacteria, parasites, chlamydia, and fungus; biomarkers, such as microRNAs, lncRNAs, and circRNAs; and some non-nucleic acid targets, including proteins, ions, and methyl groups. Meanwhile, we highlight the working principles of some novel Cas13a-based detection methods, including the Specific High-Sensitivity Enzymatic Reporter UnLOCKing (SHERLOCK) and its improved versions, Cas13a-based nucleic acid amplification-free biosensors, and Cas13a-based biosensors for non-nucleic acid target detection. Finally, we focus on some issues that need to be solved and the development prospects of the CRISPR/Cas13a system.

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“…Dual signal amplification by NASAB and Cas13a reporting endowed a high-sensitivity down to 1 CFU and 1% viable Salmonella (Fig. 7C) [110].…”

Section: Bacterial Infection Diagnosismentioning

confidence: 99%

CRISPR-based nucleic acid diagnostics for pathogens

Yang

Zhang

Teng

et al. 2022

Preprint

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Pathogenic infections remain the primary threat for human health, especially the global COVID-19 pandemic. It is important to develop rapid, sensitive and multiplexed tools for detecting pathogens and their mutations, particularly the tailor-made strategies for point-of-care diagnosis allowing for use in resource-constrained settings. The rapidly evolving CRISPR/Cas systems have provided a powerful toolbox for pathogenic diagnostics via nucleic acid tests. In this review, we first describe the resultant promising class 2 (single, multidomain effector) and recently explored class 1 (multisubunit effector complexes) CRISPR tools. We present the diverse engineering nucleic acid diagnostics based on CRISPR/Cas systems for pathogenic viruses, bacteria and fungi, and highlight the application for detecting viral variants and drug-resistant bacteria enabled by CRISPR-based mutation profiling. Finally, we discuss the challenges such as the development of preamplification-free diagnostic assays and present the emerging CRISPR systems and CRISPR cascade that potentially enable multiplexed and preamplification-free pathogenic diagnostics.

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Isothermal RNA Amplification for the Detection of Viable Pathogenic Bacteria to Estimate the Salmonella Virulence for Causing Enteritis

Cited by 40 publications

References 37 publications

CRISPR‐Cas‐based detection for food safety problems: Current status, challenges, and opportunities

CRISPR‐Cas‐based detection for food safety problems: Current status, challenges, and opportunities

CRISPR-Cas13a system: A novel tool for molecular diagnostics

CRISPR-based nucleic acid diagnostics for pathogens

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