CRISPR/Cas12a based fluorescence-enhanced lateral flow biosensor for detection of Staphylococcus aureus

Zhou, Baoqing; Ye, Qinghua; Li, Fan; Xiang, Xinran; Shang, Yuting; Wang, Chu‐Fang; Shao, Yanna; Xue, L.; Zhang, Jumei; Wang, Juan; Ding, Yu; Chen, Moutong; Wu, Qingping

doi:10.1016/j.snb.2021.130906

Cited by 76 publications

(39 citation statements)

References 35 publications

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“…The method had a detection limit of 67 CFU/mL in pure culture and 73 CFU/g in shrimp. It was more sensitive than previously reported methods, which exhibited a sensitivity of 1.35 × 10 3 CFU/mL for Listeria monocytogenes in grass carp [ 33 ], 5.4 × 10 2 CFU/mL for Staphylococcus aureus in pure culture [ 34 ], and 2 × 10 3 CFU/mL for Salmonella in powered infant formula milk [ 35 ]. It was reported that the LOD of the method using only real-time RPA for V. parahaemolyticus detection was 1.02 × 10 2 copies/reaction, but for artificially contaminated samples with different bacteria concentrations, the LODs were 4, 1, and 7 CFU/25 g in oyster sauce, codfish and sleeve-fish, respectively, after enrichment for 6 h [ 36 ].…”

Section: Discussionmentioning

confidence: 99%

CE–RAA–CRISPR Assay: A Rapid and Sensitive Method for Detecting Vibrio parahaemolyticus in Seafood

Cao²,

Zhang

et al. 2022

Foods

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Vibrio parahaemolyticus is one of the major pathogenic Vibrio species that contaminate seafood. Rapid and accurate detection is crucial for avoiding foodborne diseases caused by pathogens and is important for food safety management and mariculture. In this study, we established a system that combines chemically enhanced clustered regularly interspaced short palindromic repeats (CRISPR) and recombinase-aided amplification (RAA) (CE–RAA–CRISPR) for detecting V. parahaemolyticus in seafood. The method combines RAA with CRISPR-associated protein 12a (Cas12a) for rapid detection in a one-pot reaction, effectively reducing the risk of aerosol contamination during DNA amplifier transfer. We optimized the primers for V. parahaemolyticus, determined the optimal crRNA/Cas12a ratio, and demonstrated that chemical additives (bovine serum albumin and L-proline) could enhance the detection capacity of Cas12a. The limit of detection (at optimal conditions) was as low as 6.7 × 101 CFU/mL in pure cultures and 7.3 × 101 CFU/g in shrimp. Moreover, this method exhibited no cross-reactivity with other microbial pathogens. The CE–RAA–CRISPR assay was compared with the quantitative polymerase chain reaction assay using actual food samples, and it showed 100% diagnostic agreement.

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Section: Discussionmentioning

confidence: 99%

CE–RAA–CRISPR Assay: A Rapid and Sensitive Method for Detecting Vibrio parahaemolyticus in Seafood

Cao²,

Zhang

et al. 2022

Foods

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show abstract

“…CRISPR/Cas‐RAA‐based LFB (CRA‐LFB) was established in conjunction with functionalized quantum dots to detect S. aureus within 70 min and detection limit of genomic DNA and S. aureus were 75 aM and 5.4 × 10 2 CFU/mL, respectively. The method presents low‐cost, simple, and sensitive characteristics and excellent practical performance in natural food samples (Zhou et al., 2022). The one‐tube RPA‐CRISPR/Cas12a platform visualized by lateral flow test strips (LFS) was able to detect MRSA within 20 min.…”

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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“…The detection limit for viable S. aureus was 10 2 CFU/ml under 3 h enrichment and 10 1 CFU/ml under 6 h enrichment in artificially contaminated milk, respectively. Zhou et al (2022) reported a novel CRISPR/Cas12a-based fluorescence enhanced lateral flow biosensor (LFB) in conjunction with functionalized quantum dots, combined with RAA, to establish low-cost, simple, and sensitive detection of S. aureus , namely, CRISPR/Cas-recombinase-assisted amplification-based LFB (CRA-LFB). The limit of detection was as low as 75 aM of genomic DNA, and 5.4 × 10 2 CFU/ml of S. aureus in pure cultures were detected.…”

Section: Molecular Biology Detection Technologymentioning

confidence: 99%

Research progress on detection techniques for point-of-care testing of foodborne pathogens

Zhao

Huang

et al. 2022

Front. Bioeng. Biotechnol.

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The global burden of foodborne disease is enormous and foodborne pathogens are the leading cause of human illnesses. The detection of foodborne pathogenic bacteria has become a research hotspot in recent years. Rapid detection methods based on immunoassay, molecular biology, microfluidic chip, metabolism, biosensor, and mass spectrometry have developed rapidly and become the main methods for the detection of foodborne pathogens. This study reviewed a variety of rapid detection methods in recent years. The research advances are introduced based on the above technical methods for the rapid detection of foodborne pathogenic bacteria. The study also discusses the limitations of existing methods and their advantages and future development direction, to form an overall understanding of the detection methods, and for point-of-care testing (POCT) applications to accurately and rapidly diagnose and control diseases.

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CRISPR/Cas12a based fluorescence-enhanced lateral flow biosensor for detection of Staphylococcus aureus

Cited by 76 publications

References 35 publications

CE–RAA–CRISPR Assay: A Rapid and Sensitive Method for Detecting Vibrio parahaemolyticus in Seafood

CE–RAA–CRISPR Assay: A Rapid and Sensitive Method for Detecting Vibrio parahaemolyticus in Seafood

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

Research progress on detection techniques for point-of-care testing of foodborne pathogens

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