Establishment of a rapid method for skipjack tuna (Katsuwonus pelamis) authentication using molecular beacons in loop-mediated isothermal amplification

Xu, Wenjie; Li, Qiuping; Xue, Hanyue; Fei, Yanjin; Cui, Xiaowen; Cao, Min; Xiong, Xianrong; Yang, Ying; Wang, Libin

doi:10.1016/j.foodchem.2022.132365

Cited by 18 publications

(13 citation statements)

References 38 publications

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“…18 However, the common disadvantage of the mentioned methods is the challenge to distinguish false positives caused by non-specific amplification generated from primer dimers. 19 Moreover, the high risk of carryover contamination by the uncapping operation has also been widely recognized. 20 The main advantages of the developed method in the present work are rapid determination (about 30 min), simple operation (closed-tube format), high specificity (self-quenched probe), equipment independence (only the water bath) and visible results (naked-eye inspection).…”

Section: Methods Validation On Commercial Productsmentioning

confidence: 99%

“…Until now, the LAMP assay for rapid identification of Atlantic cod has been reported based on short fragments of mitochondrial cytochrome b (cytb) gene, where the results were obtained using real‐time amplification curve, 17 pre‐addition of pH indicator, 2 post‐addition of propidium iodide and agarose gel electrophoresis 18 . However, the common disadvantage of the mentioned methods is the challenge to distinguish false positive results caused by non‐specific amplification generated from primer dimers 19 . Moreover, the high risk of carryover contamination by the uncapping operation has also been widely recognized 20 …”

Section: Introductionmentioning

confidence: 99%

“…To achieve better specificity, target‐specific detection of LAMP amplicons can be fulfilled through the use of target‐specific probes or modified primers as the biorecognition elements 16 . For instance, the target‐specific dual‐labeled molecular beacon has been shown to be selective in the discrimination of skipjack tuna ( Katsuwonus pelamis ) 19 . Recently, a simple and cost‐effective approach based on photo‐induced electron transfer, whereby the fluorophore's quenching/dequenching occurs in the absence of a dedicated quencher, has also proved its versatility and adaptability 21 .…”

Section: Introductionmentioning

confidence: 99%

“…18 However, the common disadvantage of the mentioned methods is the challenge to distinguish false positive results caused by non-specific amplification generated from primer dimers. 19 Moreover, the high risk of carryover contamination by the uncapping operation has also been widely recognized. 20 To achieve better specificity, target-specific detection of LAMP amplicons can be fulfilled through the use of target-specific probes or modified primers as the biorecognition elements.…”

Section: Introductionmentioning

confidence: 99%

“…16 For instance, the target-specific dual-labeled molecular beacon has been shown to be selective in the discrimination of skipjack tuna (Katsuwonus pelamis). 19 Recently, a simple and cost-effective approach based on photo-induced electron transfer, whereby the fluorophore's quenching/dequenching occurs in the absence of a dedicated quencher, has also proved its versatility and adaptability. 21 Moreover, its potential to detect single-nucleotide polymorphisms of multiple genes has also been validated.…”

Section: Introductionmentioning

confidence: 99%

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Closed‐tube visual detection of Atlantic cod (Gadus morhua) using self‐quenched primer coupled with a designed loop‐mediated isothermal amplification vessel

et al. 2023

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BACKGROUND Species adulteration has been widely revealed around the world, and the possible reasons include declining stocks in most source areas of the world, poor transparency in the global supply chain, and difficulty in distinguishing the features of processed products. The present work selected Atlantic cod (Gadus morhua) as a case study, and developed a novel loop‐mediated isothermal amplification (LAMP) assay for Atlantic cod authentication, where a self‐quenched primer and a newly designed reaction vessel were used to realize the endpoint visual detection of the target‐specific products. RESULTS A novel LAMP primer set was designed for Atlantic cod, and the inner primer BIP was selected to label the self‐quenched fluorogenic element. The fluorophore's dequenching only occurred along with LAMP elongation for the target species. No fluorescence could be observed with both single‐stranded DNA and partially complementary dsDNA of the non‐target species. With the novel reaction vessel, both amplification as well as detection were operated in an enclosed device, and visual differentiation of Atlantic cod, negative control, and false positive generated from primer dimers was achieved. The novel assay has proved its specificity and applicability, and could detect as little as 1 pg of Atlantic cod DNA. Moreover, Atlantic cod adulteration as low as 10% could be detected in haddock (Melanogrammus aeglefinus), and no cross‐reactivity was observed. CONCLUSION The established assay could be a useful tool to detect mislabeling incidents involving Atlantic cod considering the advantages of speed, simplicity and accuracy. © 2023 Society of Chemical Industry.

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Section: Methods Validation On Commercial Productsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Closed‐tube visual detection of Atlantic cod (Gadus morhua) using self‐quenched primer coupled with a designed loop‐mediated isothermal amplification vessel

et al. 2023

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Evaluation of reverse transcription loop‐mediated isothermal amplification assay for the detection of severe fever with thrombocytopenia syndrome in clinical laboratories: A single‐center study

Tian,

Zhang,

Geng

et al. 2023

Journal of Medical Virology

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Severe fever with thrombocytopenia syndrome (SFTS) is an acute infectious disease prevalent in East Asia with a high mortality rate (5%–30%). Reverse transcription loop‐mediated isothermal amplification (RT‐LAMP), a rapid nucleic acid‐based diagnostic technique, is a useful alternative for the clinical diagnosis of SFTS, particularly in resource‐limited hospitals or rural clinics in SFTS virus‐endemic regions. However, the actual clinical sensitivity and specificity of RT‐LAMP remain unclear. This study evaluated the field application of RT‐LAMP. This prospective field study included 130 patients with laboratory‐confirmed SFTS from Yantai, Shandong Province, China. Two sets of RT‐LAMP primers were validated, and one set of RT‐LAMP assays was optimized for field detection. Nucleic acids of serially collected serum/plasma samples were identified using quantitative reverse transcription polymerase chain reaction (RT‐qPCR) and RT‐LAMP. In laboratory tests, we optimized the detection time of primer set 2 for the RT‐LAMP to 60 min. Notably, the onsite testing of 279 plasma samples from patients with SFTS revealed that the sensitivity and specificity of the test were 81.9% and 96.3%, respectively. We also analyzed samples with different durations of the disease, and our study showed that the sensitivity of RT‐LAMP detection at the beginning of admission was 89.92%. Univariate analysis showed that the detection rate of RT‐LAMP was similar to that of RT‐qPCR in the first 5 days of the disease course and was lower than that of RT‐qPCR on Days 6 and 14–15 of the disease course. The positive detection rate in patients aged ≥ 65 years was significantly higher than that in younger age groups. RT‐LAMP is a simple, suitable, and rapid clinical detection method of SFTS onsite screening. It is more suitable for screening patients in the early stages of the disease and analyzing samples obtained from patients aged ≥ 65 years before the 6th day of the disease course.

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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

Food Anal. Methods

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A methodology with rapidity and speci city is of great signi cance 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 ampli cation (CPA) detection method is the possibility of detecting false-positive signals derived from opening tube lids or non-speci c ampli cation, and molecular beacon was rstly employed to solve aforementioned problems. The reaction system was optimized and the results showed that the MB-CPA method was highly speci c 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% speci city 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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Establishment of a rapid method for skipjack tuna (Katsuwonus pelamis) authentication using molecular beacons in loop-mediated isothermal amplification

Cited by 18 publications

References 38 publications

Closed‐tube visual detection of Atlantic cod (Gadus morhua) using self‐quenched primer coupled with a designed loop‐mediated isothermal amplification vessel

Closed‐tube visual detection of Atlantic cod (Gadus morhua) using self‐quenched primer coupled with a designed loop‐mediated isothermal amplification vessel

Evaluation of reverse transcription loop‐mediated isothermal amplification assay for the detection of severe fever with thrombocytopenia syndrome in clinical laboratories: A single‐center study

A Novel Fluorescence cross-priming Amplification Based on Universal Molecular Beacon for Rapid and Specific Detection of Salmonella enterica in food Samples

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