Development of isothermal amplification methods for rapid and sensitive detection of heat-labile enterotoxin producing Escherichia coli

Liu, Wenxin; Yuan, Chaowen; Zhang, Liguo; Feng, Yufei

doi:10.1016/j.mimet.2019.04.010

Cited by 13 publications

(10 citation statements)

References 22 publications

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“…In 2016, CRISPR-Cas systems were first developed to identify nucleic acids for molecular diagnostics (33). Demand for instrument-free nucleic acid detection technologies has driven the development of multiple techniques for isothermal amplification (34,35). However, common approaches for isothermal amplification, such as recombinase polymerase amplification (RPA) (36), require optimization and cannot on November 25, 2020 by guest http://jcm.asm.org/ Downloaded from typically discriminate between single-base-pair differences in target sequences, a distinction that can have important consequences for pathogenicity (37)(38)(39).…”

Section: Detection Of Nucleic Acids By Crispr-cas Systemsmentioning

confidence: 99%

SHERLOCK and DETECTR: CRISPR-Cas Systems as Potential Rapid Diagnostic Tools for Emerging Infectious Diseases

2021

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Infectious diseases are one of the most intimidating threats to human race, responsible for an immense burden of disabilities and deaths. Rapid diagnosis and treatment of infectious diseases is a better understanding of its pathogenesis. According to WHO, the ideal approach for detecting foreign pathogens should be rapid, specific, sensitive, instrument-free and cost-effective. Nucleic acid pathogen detection methods, typically PCR have numerous limitations, such as highly sophisticated equipments, reagents, and trained personnel rely on well-established laboratories beside time-consuming. Thus, there is a crucial need to develop novel nucleic acid detection tools with rapid, specific, sensitive, and cost-effective, particularly ones that can be used for versatile point-of-care diagnostic applications. Two new methods exploit on unpredicted in vitro properties CRISPR-Cas effectors, turning activated nucleases into basic amplifiers of a specific nucleic-acid binding event. These effectors are attached with a diversity of reporters and utilized in tandem with present of isothermal amplification approaches to create sensitive identification in multiple field deployable formats. Although still in their beginning, yet SHERLOCK and DETECTR technologies are potential methods for rapid detection and identification of infectious disease, with ultra-sensitive tests that don't require a lot of complicated processing. This review described SHERLOCK and DETECTR technologies beside their properties, functions, and perspectives to become the ultimate diagnostic tools for diagnosing infectious diseases and curbing disease outbreaks.

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Section: Detection Of Nucleic Acids By Crispr-cas Systemsmentioning

confidence: 99%

SHERLOCK and DETECTR: CRISPR-Cas Systems as Potential Rapid Diagnostic Tools for Emerging Infectious Diseases

2021

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“…In recent studies, Liu et al reported the development of Loop-mediated isothermal amplification (LAMP), cross-priming amplification (CPA) and isothermal multiple self-matching initiated amplification (IMSA) to detect heat-labile enterotoxin-producing Escherichia coli. The three isothermal amplification assays demonstrated high specificity and sensitivity when detecting heat-labile enterotoxin-producing E. coli in clinical samples [24,28]. Additionally, research by Yang, et al also confirmed that IMSA demonstrated higher sensitivity than LAMP or CPA assays [19].…”

Section: Discussionmentioning

confidence: 83%

“…Multiple reports have been published indicating that CPA technology has been successfully used as a specific and sensitive method for the detection of Mycobacterium tuberculosis, Enterobacter sakazakii and heat labile enterotoxin of Escherichia coli etc. [20,23,24].…”

Section: Introductionmentioning

confidence: 99%

Establishment and application of isothermal amplification techniques for the detection of heat-stable I enterotoxin of enterotoxigenic Escherichia coli

et al. 2020

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The aim of this study was to establish isothermal multiple self-matching initiated amplification (IMSA) and cross-priming amplification (CPA) methods to detect heat-stable I enterotoxin (STa)-producing enterotoxigenic Escherichia coli (ETEC). To avoid cross-contamination of aerosols, a closed independent isothermal amplification tube (IAT) was used to perform the assays. Optimal amplification conditions for IMSA and CPA were selected for specificity and sensitivity, respectively, and for clinical relevance. Both IMSA and CPA assays could specifically recognize all 3-STa positive strains in which they fluoresced green under UV light, but not in the 11 non-STa strains. The results of the sensitivity analysis indicated that the detection limit of the IMSA assay was 1.5 ×10 2 CFU, comparable to real-time PCR, but 10-fold more sensitive than CPA and LAMP. Further evaluation of the detection methods of swine diarrhea samples demonstrated that both could successfully identify the DNA of STa-producing ETEC in clinical specimens, consistent with LAMP and qPCR methods. The results demonstrated that the IMSA and CPA methods had high specificity and sensitivity with rapid detection of ETEC, so having great potential in clinical applications.

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“…[32] Demand for instrument-free nucleic acid detection technologies has driven the development of multiple techniques for isothermal amplification. [33,34] However, common approaches for isothermal amplification, such as recombinase polymerase amplification (RPA) [35], require optimization and cannot typically discriminate between single-base-pair differences in target sequences, a distinction that can have important consequences for pathogenicity [36][37][38]. Recently, enzymes from CRISPR-Cas systems have been adapted for the specific, rapid, sensitive, and portable detecting of nucleic acids.…”

Section: Detection Of Nucleic Acids By Crispr-casmentioning

confidence: 99%

<strong>SHERLOCK and DETECTR: CRISPR-Cas Systems as Potential Rapid Diagnostic Tools for Emerging Infectious Diseases and Cancer-Associated Mutations</strong>

Mustafa¹,

Makhawi²

2020

Preprint

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Sensitive and precise nucleic acid detection is critical for clinical diagnostics and biotechnological advancements. Diagnostic in infectious disease field is very unique from diagnosing any other disease, that is time is of the essence; in outbreaks people die even with each passing hour in some cases, if the correct diagnosis wasn't make; for example Zika in particularly is a very challenging virus to diagnose, because it's in very few numbers of copies in the infected person, so it need high sensitive diagnostic approach to spot it, In particular, the advanced tools SHERLOCKv2 and DETECTR, give almost an immediate detection of attomolar amounts of pathogenic nucleic acids with specificity similar to that of PCR but with slight technical settings and that will guide the correct intervention for the patient. SHERLOCKv2 and DETECTR technologies are game changers for our ability to identify infectious disease and rapid detection of tumor DNA or cancer-related viruses with ultra-sensitive tests that don’t require a lot of complicated processing to go through. In this paper, we will review cutting-edge infectious disease diagnosis by CRISPR-Cas systems.

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Development of isothermal amplification methods for rapid and sensitive detection of heat-labile enterotoxin producing Escherichia coli

Cited by 13 publications

References 22 publications

SHERLOCK and DETECTR: CRISPR-Cas Systems as Potential Rapid Diagnostic Tools for Emerging Infectious Diseases

SHERLOCK and DETECTR: CRISPR-Cas Systems as Potential Rapid Diagnostic Tools for Emerging Infectious Diseases

Establishment and application of isothermal amplification techniques for the detection of heat-stable I enterotoxin of enterotoxigenic Escherichia coli

<strong>SHERLOCK and DETECTR: CRISPR-Cas Systems as Potential Rapid Diagnostic Tools for Emerging Infectious Diseases and Cancer-Associated Mutations</strong>

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