A rapid and colorimetric loop-mediated isothermal amplification (LAMP) based on HRP-mimicking molecular beacon for the detection of major 6 Listeria species in enoki mushroom

Lee, Jeong Eun; Kim, Sol-A; Mun, Hyoyoung; Kim, Se-Ri; Ha, Kwang-Soo; Shim, Won‐Bo

doi:10.1016/j.foodcont.2021.108569

Cited by 14 publications

(5 citation statements)

References 46 publications

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“…Table S2 ( Supplementary Materials ) outlines important features of LAMP protocols compiled through a systematic literature review pertaining to the highly specific L. monocytogenes detection. The probit estimated LOD 95 (0.5 CFU mL −1 ) was within the same order of magnitude as those values documented by Wachiralurpan et al [ 56 ] and Lee et al [ 15 ], which developed LAMP protocols with the capability of detecting as low as 0.3–3 and 1 CFU mL −1 , respectively. The comparison with other detection thresholds gathered from the available literature ( Table S2 ) highlighted the superior analytical performance of the current method, which displayed a LOD 95 value 6- to 20,000-fold lower [ 14 , 15 , 17 , 57 , 58 , 59 , 60 ].…”

Section: Resultssupporting

confidence: 84%

“…Moreover, LAMP has proven superior analytical sensitivity, vanquishing PCR-based systems performance [ 14 ]. Beyond the well-documented outstanding LAMP performance [ 14 , 15 , 16 , 17 ], the versatility of the endpoint readout (e.g., turbidimetry, colourimetry, electrochemistry, fluorescence) is an appealing trait, contributing to the practical implementation in resource-scarce laboratories/facilities, or for field purposes.…”

Section: Introductionmentioning

confidence: 99%

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Development of a Novel Phagomagnetic-Assisted Isothermal DNA Amplification System for Endpoint Electrochemical Detection of Listeria monocytogenes

et al. 2023

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The hitherto implemented Listeria monocytogenes detection techniques are cumbersome or require expensive non-portable instrumentation, hindering their transposition into on-time surveillance systems. The current work proposes a novel integrated system resorting to loop-mediated isothermal amplification (LAMP), assisted by a bacteriophage P100–magnetic platform, coupled to an endpoint electrochemical technique, towards L. monocytogenes expeditious detection. Molybdophosphate-based optimization of the bacterial phagomagnetic separation protocol allowed the determination of the optimal parameters for its execution (pH 7, 25 °C, 32 µg of magnetic particles; 60.6% of specific capture efficiency). The novel LAMP method targeting prfA was highly specific, accomplishing 100% inclusivity (for 61 L. monocytogenes strains) and 100% exclusivity (towards 42 non-target Gram-positive and Gram-negative bacteria). As a proof-of-concept, the developed scheme was successfully validated in pasteurized milk spiked with L. monocytogenes. The phagomagnetic-based approach succeeded in the selective bacterial capture and ensuing lysis, triggering Listeria DNA leakage, which was efficiently LAMP amplified. Methylene blue-based electrochemical detection of LAMP amplicons was accomplished in 20 min with remarkable analytical sensitivity (1 CFU mL−1). Hence, the combined system presented an outstanding performance and robustness, providing a 2.5 h-swift, portable, cost-efficient detection scheme for decentralized on-field application.

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Section: Resultssupporting

confidence: 84%

Section: Introductionmentioning

confidence: 99%

Development of a Novel Phagomagnetic-Assisted Isothermal DNA Amplification System for Endpoint Electrochemical Detection of Listeria monocytogenes

et al. 2023

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“…In this study, molecular beacon containing target invA sequence and complimentary sequence that can intercalate to CPA amplicons of S. enterica were used for the development real-time CPA. As we know, only the loop position is single stranded and suits for the beacon to bind in the LAMP reaction resulted from its speci c structure (Lee, et al, 2022). However, the products forms of CPA are different, and there are many regions that we can design the probes.…”

Section: Mb Probes Designing and Optimizationmentioning

confidence: 99%

A novel fluorescence cross-priming amplification based on universal molecular beacon for rapid and specific detection of Salmonella enterica in food samples

Hou,

Du,

Liu

et al. 2024

Preprint

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A methodology with rapidity and specificity is of great significance for effectively control and management of disease epidemics caused by Salmonella enterica as it has presented an obvious threat to food safety and public health worldwide. One major drawback to the traditional cross-priming amplification (CPA) detection method is the possibility of detecting false-positive signals derived from opening tube lids or non-specific amplification, and molecular beacon was firstly employed to solve aforementioned problems. The reaction system was optimized and the results showed that the MB-CPA method was highly specific for detection of S. enterica. The LOD of established assay was found to be 10 CFU/mL, 40 CFU/mL, 4 CFU/mL in pure culture, chicken sample without and with 6 h enrichment, respectively. And the LOD of MB-CPA was 10 times higher than that of real-time PCR. An application of MB-CPA assay was conducted with 78 naturally contaminated food samples to test its practicality. After an enrichment step at 37℃ for 6h, the results showed 100% sensitivity and 100% specificity compared with standard culture-based method. Considering its rapidity, user-friendliness, cost-effectiveness, this MB-CPA assay will aid in the broader application in food industry for the detection of S. enterica in small or resource-limited food testing laboratories.

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“…To address this constraint, the conjunction of nucleic acid amplication technologies and HGD has been employed to achieve substantial signal amplication, elevating the detection capabilities by orders of magnitude. Various nucleic acid amplication techniques are being integrated with HGD for detection purposes, such as Polymerase Chain Reaction (PCR), [32][33][34][35][36][37] Loop-Mediated Isothermal Amplication (LAMP), [38][39][40][41][42][43][44] and Recombinase Polymerase Amplication (RPA). [45][46][47][48] These methods are able to amplify the signal to be detected.…”

Section: Amplication Of Target and Detectionmentioning

confidence: 99%

“…55 On the advancement of biosensors for the purpose of bacterial detection, Lee and coworkers developed a detection technique for Escherichia coli O157:H7 and Listeria utilizing a molecular beacon and LAMP. 38,39 The molecular beacon contains a sequence of the G-quadruplex. When it combines with the LAMP amplicon, it cannot form HGD.…”

Section: Amplication Of Target and Detectionmentioning

confidence: 99%

Recent progress on DNAzyme-based biosensors for pathogen detection

Liu,

Yuan,

Xiao

2024

Anal. Methods

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A rapid and colorimetric loop-mediated isothermal amplification (LAMP) based on HRP-mimicking molecular beacon for the detection of major 6 Listeria species in enoki mushroom

Cited by 14 publications

References 46 publications

Development of a Novel Phagomagnetic-Assisted Isothermal DNA Amplification System for Endpoint Electrochemical Detection of Listeria monocytogenes

Development of a Novel Phagomagnetic-Assisted Isothermal DNA Amplification System for Endpoint Electrochemical Detection of Listeria monocytogenes

A novel fluorescence cross-priming amplification based on universal molecular beacon for rapid and specific detection of Salmonella enterica in food samples

Recent progress on DNAzyme-based biosensors for pathogen detection

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