Development of Recombinase-Aided Amplification (RAA)-Exo-Probe and RAA-CRISPR/Cas12a Assays for Rapid Detection of <i>Campylobacter jejuni</i> in Food Samples

Zhi, Shuai; Shen, Jinling; Li, Xingang; Jiang, Yuan; Xue, Junxin; Fang, Taisong; Xu, Jin; Wang, Xuan; Cao, Yuhao; Yang, Danting; Yao, Zhiyuan; Yu, Daniel

doi:10.1021/acs.jafc.2c02581

Cited by 18 publications

(13 citation statements)

References 40 publications

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“…These results confirmed the superior selectivity of our strategy toward MPXV, which is derived from the outstanding specificity of RAA amplification and Cas12a recognition. 47-49…”

Section: Resultsmentioning

confidence: 99%

CRISPR/Cas12a-mediated ultrasensitive and on-site monkeypox viral testing

Zhao

Wang

et al. 2022

Preprint

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The unexpected transmission of monkeypox virus (MPXV) from Central and West Africa to previously non-endemic locations is triggering a global panic. The ultrasensitive, rapid, and specific detection of MPXV is crucial for controlling its spreading, while such technology has rarely been reported. Herein, we proposed an MPXV assay combining recombinase-aided amplification (RAA) and CRISPR/Cas12a for the first time. This assay targeted MPXV F3L gene and yielded a low detection limit (LOD) of 101 copies/μL. Deriving from the high specificity nature of RAA and CRISPR/Cas12a, through rational optimizations of probes and conditions, this assay showed high selectivity that could distinguish MPXV from other orthopox viruses and current high-profile viruses. To facilitate on-site screening of potential MPXV carriers, a kit integrating lateral flow strips was developed, enabling naked-eye MPXV detection with a LOD of 104 copies/μL. Our RAA-Cas12a-MPXV assay was able to detect MPXV without the need for sophisticated operation and expensive equipment. We envision that this RAA-Cas12a-MPXV assay can be deployed in emerging viral outbreaks for on-site surveillance of MPXV.

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“…These results confirmed the superior selectivity of our strategy toward MPXV, which is derived from the outstanding specificity of RAA amplification and Cas12a recognition. 47-49…”

Section: Resultsmentioning

confidence: 99%

CRISPR/Cas12a-mediated ultrasensitive and on-site monkeypox viral testing

Zhao

Wang

et al. 2022

Preprint

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“…In addition, real-time RT-RAA amplification can also be combined with microfluidic chip technology for multipathogen detection [ 25 ]. In addition, several studies have shown that the combination of RAA technology and the CRISPR system can achieve ultrahigh sensitivity and specific detection of single DNA or RNA molecules [ 26 , 27 , 28 ]. Because RAA technology is specific, sensitive, rapid, and easy to operate, it has great application prospects in the early diagnosis of animal diseases, immediate detection, and import- and export-related quarantine.…”

Section: Discussionmentioning

confidence: 99%

Real-Time Reverse Transcription Recombinase-Aided Amplification Assay for Rapid Amplification of the N Gene of SARS-CoV-2

Tu¹,

Zhang

Zhang³

et al. 2022

IJMS

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COVID-19 was officially declared a global pandemic disease on 11 March 2020, with severe implications for healthcare systems, economic activity, and human life worldwide. Fast and sensitive amplification of the severe acute respiratory syndrome virus 2 (SARS-CoV-2) nucleic acids is critical for controlling the spread of this disease. Here, a real-time reverse transcription recombinase-aided amplification (RT-RAA) assay, targeting conserved positions in the nucleocapsid protein gene (N gene) of SARS-CoV-2, was successfully established for SARS-CoV-2. The assay was specific to SARS-CoV-2, and there was no cross-reaction with other important viruses. The sensitivity of the real-time RT-RAA assay was 142 copies per reaction at 95% probability. Furthermore, 100% concordance between the real-time RT-RAA and RT-qPCR assays was achieved after testing 72 clinical specimens. Further linear regression analysis indicated a significant correlation between the real-time RT-RAA and RT-qPCR assays with an R2 value of 0.8149 (p < 0.0001). In addition, the amplicons of the real-time RT-RAA assay could be directly visualized by a portable blue light instrument, making it suitable for the rapid amplification of SARS-CoV-2 in resource-limited settings. Therefore, the real-time RT-RAA method allows the specific, sensitive, simple, rapid, and reliable detection of SARS-CoV-2.

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“…The improved specificity of Cas14a makes its usage for ssDNA detection possible without the need for PAM sequence. [30] Campylobacter jejuni Fluorescence Cas12a / / 5 copies / reaction [312] fluorescence, electrochemical, electrochemiluminescence (ECL), and SERS signals) and advanced nanotechnology. Herein, the construction and principle of CRISPR/Cas systemspowered bio/nanosensors will be discussed according to the different signal transduction models.…”

Section: Cas14mentioning

confidence: 99%

CRISPR/Cas Systems‐Inspired Nano/Biosensors for Detecting Infectious Viruses and Pathogenic Bacteria

et al. 2022

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Infectious pathogens cause severe human illnesses and great deaths per year worldwide. Rapid, sensitive, and accurate detection of pathogens is of great importance for preventing infectious diseases caused by pathogens and optimizing medical healthcare systems. Inspired by a microbial defense system (i.e., CRISPR/ CRISPR-associated proteins (Cas) system, an adaptive immune system for protecting microorganisms from being attacked by invading species), a great many new biosensors have been successfully developed and widely applied in the detection of infectious viruses and pathogenic bacteria. Moreover, advanced nanotechnologies have also been integrated into these biosensors to improve their detection stability, sensitivity, and accuracy. In this review, the recent advance in CRISPR/Cas systems-based nano/biosensors and their applications in the detection of infectious viruses and pathogenic bacteria are comprehensively reviewed. First of all, the categories and working principles of CRISPR/Cas systems for establishing the nano/biosensors are simply introduced. Then, the design and construction of CRISPR/Cas systems-based nano/biosensors are comprehensively discussed. In the end, attentions are focused on the applications of CRISPR/Cas systems-based nano/biosensors in the detection of infectious viruses and pathogenic bacteria. Impressively, the remaining opportunities and challenges for the further design and development of CRISPR/Cas system-based nano/biosensors and their promising applications are proposed.

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Development of Recombinase-Aided Amplification (RAA)-Exo-Probe and RAA-CRISPR/Cas12a Assays for Rapid Detection of Campylobacter jejuni in Food Samples

Cited by 18 publications

References 40 publications

CRISPR/Cas12a-mediated ultrasensitive and on-site monkeypox viral testing

CRISPR/Cas12a-mediated ultrasensitive and on-site monkeypox viral testing

Real-Time Reverse Transcription Recombinase-Aided Amplification Assay for Rapid Amplification of the N Gene of SARS-CoV-2

CRISPR/Cas Systems‐Inspired Nano/Biosensors for Detecting Infectious Viruses and Pathogenic Bacteria

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