Development of loop-mediated isothermal amplification (LAMP) assays for the rapid detection of allergic peanut in processed food

Sheu, Shyang-Chwen; Tsou, Po-Chuan; Lien, Yi-Yang; Lee, Meng-Shiou

doi:10.1016/j.foodchem.2018.02.124

Cited by 44 publications

(27 citation statements)

References 16 publications

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“…In some references, a spacer was inserted between F1c or B1c and F2 or B2 in the inner primer (FIP or BIP) [16,39]. However, the spacer was not used in this study, because it was shown that the LAMP reaction can also progress without the use of spacers [25,27]. Primer sets were designed to target the atpA, ITS1, ITS2, and the ITS1 and ITS2 gene spanning the whole 5.8S gene and to target the ORF160b (Table 3, Figures S1-S3).…”

Section: Target Gene Selection Alignments and Primer Designmentioning

confidence: 99%

“…The only equipment required is a simple thermostatically controlled water bath, heat block, or oven [18,19].With regard to the food and feed sector, mainly LAMP methods for the detection of foodborne pathogens [21,22] and genetically modified crops [23,24] are described. However, a number of reports suggest that LAMP may also be useful for the detection of allergenic foods [25][26][27][28]. LAMP amplicons can be detected by various methods, e.g., agarose gel electrophoresis (AGE) [25,27,28], real-time fluorescence detection using intercalating nucleic acid dyes [26][27][28], microfluidic chips for visual detection based on a pH indicator dye [26], visual detection using intercalating nucleic acid dyes [27], color changes of metal-sensitive indicators, such as calcein [28], and the visual or turbidimeter detection of turbidity formation [24].…”

mentioning

confidence: 99%

“…However, a number of reports suggest that LAMP may also be useful for the detection of allergenic foods [25][26][27][28]. LAMP amplicons can be detected by various methods, e.g., agarose gel electrophoresis (AGE) [25,27,28], real-time fluorescence detection using intercalating nucleic acid dyes [26][27][28], microfluidic chips for visual detection based on a pH indicator dye [26], visual detection using intercalating nucleic acid dyes [27], color changes of metal-sensitive indicators, such as calcein [28], and the visual or turbidimeter detection of turbidity formation [24]. AGE and real-time fluorescence monitoring require additional equipment for the reaction confirmation, which compromises the simplicity of LAMP and limits the field-like or on-site applications of this procedure.…”

mentioning

confidence: 99%

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The Development of Highly Specific and Sensitive Primers for the Detection of Potentially Allergenic Soybean (Glycine max) Using Loop-Mediated Isothermal Amplification Combined with Lateral Flow Dipstick (LAMP-LFD)

Allgöwer

Hartmann

Holzhauser

2020

Foods

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The soybean (Glycine max) has been recognized as a frequent elicitor of food allergy worldwide. A lack of causative immunotherapy of soybean allergy makes soybean avoidance essential. Therefore, sensitive and specific methods for soybean detection are needed to allow for soybean verification in foods. Loop-mediated isothermal amplification (LAMP) represents a rapid and simple DNA-based detection method principally suitable for field-like applications or on-site analytical screening for allergens during the manufacturing of foods. This work describes the systematic development and selection of suitable LAMP primers based on soybean multicopy genes. The chemistry applied allows for a versatile detection of amplified DNA, using either gel electrophoresis, fluorescence recording, or a simple Lateral Flow Dipstick (LFD). LAMP based on the ORF160b gene was highly specific for the soybean and may allow for a detection level equivalent to approximately 10 mg soy per kg food. Various soybean cultivars were detectable at a comparable level of sensitivity. LAMP combined with LFD-like detection facilitates a simple, highly specific and sensitive detection of the soybean without the need for expensive analytical equipment. In contrast to the majority of antibody-based methods for soybean detection, all identified primer sequences and optimized protocols are disclosed and broadly available to the community.Foods 2020, 9, 423 2 of 19 soybeans may remain indiscernible or hidden due to mislabeling or the cross contact of allergen-free food products with allergenic foods, e.g., during the food manufacturing process [7]. As part of the Allergen Bureau of Australia & New Zealand, the VITAL (Voluntary Incidental Trace Allergen Labeling) expert panel established reference doses for allergenic foods on the basis of individual clinical threshold data. Accordingly, 1 mg soy protein, equivalent to 2.5 mg soy, was modeled as the eliciting dose (ED) for an allergic reaction in 5% of the soy allergic population (ED05) [8]. Thus, 95% of soy allergic subjects are thought to be safe at a level of 2.5 mg soy. Recently, the soy protein reference dose was lowered to 0.5 mg, however, without peer-reviewed publication. Accordingly, for the detection of 1.25−2.5 mg soy in a serving size of 100 g, a method sensitivity from less than 12.5 to 25 mg soy per kg food is required to verify the absence or presence of soy at a level that likely improves food safety for the majority of soy allergic subjects. The availability of specific and sensitive methods for the detection of allergenic foods is essential for the verification of compliance with labeling requirements and the risk management of cross contact with allergenic foods, such as those based on the implementation of VITAL reference doses.Currently, the two most prominent analytical techniques for soybean detection in food are protein-based enzyme-linked immunosorbent assays (ELISA) [9,10] and nucleic acid (DNA)-based quantitative polymerase chain reaction (qPCR) assays [11,12]. Both types of methods ...

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Section: Target Gene Selection Alignments and Primer Designmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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The Development of Highly Specific and Sensitive Primers for the Detection of Potentially Allergenic Soybean (Glycine max) Using Loop-Mediated Isothermal Amplification Combined with Lateral Flow Dipstick (LAMP-LFD)

Allgöwer

Hartmann

Holzhauser

2020

Foods

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“…In 2000, Notomi et al developed a loop-mediated isothermal amplification (LAMP), which has the advantages of high sensitivity, good specificity, and simple operation (Domesle et al 2017 ; Chen et al 2017 ). The LAMP amplification principle is based on the use of 4 primers that are designed for 6 specific regions of the target gene using the strand displacement characteristics of Bst DNA polymerase (Velders et al 2018 ; Feng et al 2017 ; Zheney et al 2018 ; Sheu et al 2018 ). At certain temperatures, the 4 primers identify 6 specific regions of the target gene and continuously the process of extension replacement.…”

Section: Introductionmentioning

confidence: 99%

Loop-mediated isothermal amplification-lateral-flow dipstick (LAMP-LFD) to detect Mycoplasma ovipneumoniae

Zhang

Cao

Zhu

et al. 2019

World J Microbiol Biotechnol

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In order to establish a rapid detection method for Mycoplasma ovipneumoniae , this study used the loop-mediated isothermal amplification (LAMP) technique to carry out nucleic acid amplification and chromatographic visualization via a lateral flow dipstick (LFD) assay. The M. ovipneumoniae elongation factor TU gene ( EF-TU ) was detected using a set of specific primers designed for the EF-TU gene, and the EF-TU FIP was detected by biotin labeling, which was used in the LAMP amplification reaction. The digoxin-labeled probe specifically hybridized with LAMP products, which were visually detected by LFD. Here, we established the M. ovipneumoniae LAMP-LFD rapid detection method and tested the specificity, sensitivity, and clinical application of this method. Results showed that the optimized LAMP performed at 60 °C for 60 min, and LFD can specifically and visually detect M. ovipneumoniae with a minimum detectable concentration at 1.0 × 10 2 CFU/mL. The sensitivity of LAMP-LFD was 1000 times that of the conventional PCR detection methods, and the clinical lung tissue detection rate was 86% of 50 suspected sheep infected with M. ovipneumoniae . In conclusion, LAMP-LFD was established in this study to detect M. ovipneumoniae , a method that was highly specific, sensitive, and easy to operate, and provides a new method for the prevention and diagnosis of M. ovipneumoniae infection. Electronic supplementary material The online version of this article (10.1007/s11274-019-2601-5) contains supplementary material, which is available to authorized users.

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“…Since this analytical method is still mainly used in research laboratories, new approaches are being investigated to make it practical, such as developing a portable detection prototype based on Arduino, an open electronics micro controller board [81]. Currently, LAMP is more accurate than quantitative-PCR for the detection of meningococcal infections in children [82], gene doping therapies [83], peanut allergens in processed food [84], acute viral necrobiotic infections in scallops [85], and Staphylococcus pseudintermedius bacterial infections in canines [86].…”

Section: Loop-mediated Isothermal Amplification (Lamp)mentioning

confidence: 99%

DNA Microsystems for Biodiagnosis

2020

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Researchers are continuously making progress towards diagnosis and treatment of numerous diseases. However, there are still major issues that are presenting many challenges for current medical diagnosis. On the other hand, DNA nanotechnology has evolved significantly over the last three decades and is highly interdisciplinary. With many potential technologies derived from the field, it is natural to begin exploring and incorporating its knowledge to develop DNA microsystems for biodiagnosis in order to help address current obstacles, such as disease detection and drug resistance. Here, current challenges in disease detection are presented along with standard methods for diagnosis. Then, a brief overview of DNA nanotechnology is introduced along with its main attractive features for constructing biodiagnostic microsystems. Lastly, suggested DNA-based microsystems are discussed through proof-of-concept demonstrations with improvement strategies for standard diagnostic approaches.

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Development of loop-mediated isothermal amplification (LAMP) assays for the rapid detection of allergic peanut in processed food

Cited by 44 publications

References 16 publications

The Development of Highly Specific and Sensitive Primers for the Detection of Potentially Allergenic Soybean (Glycine max) Using Loop-Mediated Isothermal Amplification Combined with Lateral Flow Dipstick (LAMP-LFD)

The Development of Highly Specific and Sensitive Primers for the Detection of Potentially Allergenic Soybean (Glycine max) Using Loop-Mediated Isothermal Amplification Combined with Lateral Flow Dipstick (LAMP-LFD)

Loop-mediated isothermal amplification-lateral-flow dipstick (LAMP-LFD) to detect Mycoplasma ovipneumoniae

DNA Microsystems for Biodiagnosis

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