LAMP-LFD Based on Isothermal Amplification of Multicopy Gene <i>ORF160b</i>: Applicability for Highly Sensitive Low-Tech Screening of Allergenic Soybean (<i>Glycine max</i>) in Food
Soybean (<i>Glycine max</i>) allergy can be life threatening. A lack of causative immunotherapy of soybean allergy makes soybean avoidance indispensable. Detection methods are essential to verify allergen labeling and unintentional allergen cross contact during food manufacture. Here, we...
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| Format: | Article |
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MDPI AG
2020-11-01
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| Series: | Foods |
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| Online Access: | https://www.mdpi.com/2304-8158/9/12/1741 |
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| author | Stefanie M. Allgöwer Chris A. Hartmann Clarissa Lipinski Vera Mahler Stefanie Randow Elke Völker Thomas Holzhauser |
| author_facet | Stefanie M. Allgöwer Chris A. Hartmann Clarissa Lipinski Vera Mahler Stefanie Randow Elke Völker Thomas Holzhauser |
| author_sort | Stefanie M. Allgöwer |
| collection | DOAJ |
| description | Soybean (<i>Glycine max</i>) allergy can be life threatening. A lack of causative immunotherapy of soybean allergy makes soybean avoidance indispensable. Detection methods are essential to verify allergen labeling and unintentional allergen cross contact during food manufacture. Here, we aimed at evaluating our previously described primers for loop-mediated isothermal amplification (LAMP) of multicopy gene <i>ORF160b</i>, combined with a lateral flow dipstick (LFD)-like detection, for their performance of soybean detection in complex food matrices. The results were compared with those obtained using quantitative real-time Polymerase Chain Reaction (qPCR) as the current standard of DNA-based allergen detection, and antibody-based commercial lateral flow device (LFD) as the current reference of protein-based rapid allergen detection. LAMP-LFD allowed unequivocal and reproducible detection of 10 mg/kg soybean incurred in three representative matrices (boiled sausage, chocolate, instant tomato soup), while clear visibility of positive test lines of two commercial LFD tests was between 10 and 10<sup>2</sup> mg/kg and depending on the matrix. Sensitivity of soybean detection in incurred food matrices, commercial retail samples, as well as various processed soybean products was comparable between LAMP-LFD and qPCR. The DNA-based LAMP-LFD proved to be a simple and low-technology soybean detection tool, showing sensitivity and specificity that is comparable or superior to the investigated commercial protein-based LFD. |
| first_indexed | 2024-03-10T14:34:07Z |
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| id | doaj.art-89517856ace94a37afbdef047e55b098 |
| institution | Directory Open Access Journal |
| issn | 2304-8158 |
| language | English |
| last_indexed | 2024-03-10T14:34:07Z |
| publishDate | 2020-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Foods |
| spelling | doaj.art-89517856ace94a37afbdef047e55b0982023-11-20T22:22:45ZengMDPI AGFoods2304-81582020-11-01912174110.3390/foods9121741LAMP-LFD Based on Isothermal Amplification of Multicopy Gene <i>ORF160b</i>: Applicability for Highly Sensitive Low-Tech Screening of Allergenic Soybean (<i>Glycine max</i>) in FoodStefanie M. Allgöwer0Chris A. Hartmann1Clarissa Lipinski2Vera Mahler3Stefanie Randow4Elke Völker5Thomas Holzhauser6Division of Allergology, Paul-Ehrlich-Institut (PEI), D-63225 Langen, GermanyDivision of Allergology, Paul-Ehrlich-Institut (PEI), D-63225 Langen, GermanyDivision of Allergology, Paul-Ehrlich-Institut (PEI), D-63225 Langen, GermanyDivision of Allergology, Paul-Ehrlich-Institut (PEI), D-63225 Langen, GermanyDivision of Allergology, Paul-Ehrlich-Institut (PEI), D-63225 Langen, GermanyDivision of Allergology, Paul-Ehrlich-Institut (PEI), D-63225 Langen, GermanyDivision of Allergology, Paul-Ehrlich-Institut (PEI), D-63225 Langen, GermanySoybean (<i>Glycine max</i>) allergy can be life threatening. A lack of causative immunotherapy of soybean allergy makes soybean avoidance indispensable. Detection methods are essential to verify allergen labeling and unintentional allergen cross contact during food manufacture. Here, we aimed at evaluating our previously described primers for loop-mediated isothermal amplification (LAMP) of multicopy gene <i>ORF160b</i>, combined with a lateral flow dipstick (LFD)-like detection, for their performance of soybean detection in complex food matrices. The results were compared with those obtained using quantitative real-time Polymerase Chain Reaction (qPCR) as the current standard of DNA-based allergen detection, and antibody-based commercial lateral flow device (LFD) as the current reference of protein-based rapid allergen detection. LAMP-LFD allowed unequivocal and reproducible detection of 10 mg/kg soybean incurred in three representative matrices (boiled sausage, chocolate, instant tomato soup), while clear visibility of positive test lines of two commercial LFD tests was between 10 and 10<sup>2</sup> mg/kg and depending on the matrix. Sensitivity of soybean detection in incurred food matrices, commercial retail samples, as well as various processed soybean products was comparable between LAMP-LFD and qPCR. The DNA-based LAMP-LFD proved to be a simple and low-technology soybean detection tool, showing sensitivity and specificity that is comparable or superior to the investigated commercial protein-based LFD.https://www.mdpi.com/2304-8158/9/12/1741multicopy generapid testloop-mediated isothermal amplification (LAMP)lateral flow dipstick/device (LFD)qPCRfood allergy |
| spellingShingle | Stefanie M. Allgöwer Chris A. Hartmann Clarissa Lipinski Vera Mahler Stefanie Randow Elke Völker Thomas Holzhauser LAMP-LFD Based on Isothermal Amplification of Multicopy Gene <i>ORF160b</i>: Applicability for Highly Sensitive Low-Tech Screening of Allergenic Soybean (<i>Glycine max</i>) in Food Foods multicopy gene rapid test loop-mediated isothermal amplification (LAMP) lateral flow dipstick/device (LFD) qPCR food allergy |
| title | LAMP-LFD Based on Isothermal Amplification of Multicopy Gene <i>ORF160b</i>: Applicability for Highly Sensitive Low-Tech Screening of Allergenic Soybean (<i>Glycine max</i>) in Food |
| title_full | LAMP-LFD Based on Isothermal Amplification of Multicopy Gene <i>ORF160b</i>: Applicability for Highly Sensitive Low-Tech Screening of Allergenic Soybean (<i>Glycine max</i>) in Food |
| title_fullStr | LAMP-LFD Based on Isothermal Amplification of Multicopy Gene <i>ORF160b</i>: Applicability for Highly Sensitive Low-Tech Screening of Allergenic Soybean (<i>Glycine max</i>) in Food |
| title_full_unstemmed | LAMP-LFD Based on Isothermal Amplification of Multicopy Gene <i>ORF160b</i>: Applicability for Highly Sensitive Low-Tech Screening of Allergenic Soybean (<i>Glycine max</i>) in Food |
| title_short | LAMP-LFD Based on Isothermal Amplification of Multicopy Gene <i>ORF160b</i>: Applicability for Highly Sensitive Low-Tech Screening of Allergenic Soybean (<i>Glycine max</i>) in Food |
| title_sort | lamp lfd based on isothermal amplification of multicopy gene i orf160b i applicability for highly sensitive low tech screening of allergenic soybean i glycine max i in food |
| topic | multicopy gene rapid test loop-mediated isothermal amplification (LAMP) lateral flow dipstick/device (LFD) qPCR food allergy |
| url | https://www.mdpi.com/2304-8158/9/12/1741 |
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