REASSURED Test System for Food Control—Preparation of LAMP Reaction Mixtures for In-Field Identification of Plant and Animal Species
Source abstract
Abstract
The potential of loop-mediated isothermal amplification (LAMP) assays for species identification in medical diagnostics, food safety, and authentication is indisputable. The challenge in commercialization of such DNA-based rapid test methods for resource-limited settings is the on-site availability of the required reagents and an intuitive read-out system. In this work, reaction mixtures for LAMP assays for the detection of animal (plaice) and plant food (safflower) were lyophilized and stored at room temperature for up to 24 weeks, demonstrating that refrigeration of reagents after lyophilization is not absolutely necessary. During this period, reaction mixtures were stable and the polymerase showed no loss of activity. In addition, mobile testing, including DNA isolation, using the lyophilized LAMP reaction mixtures and a handheld fluorescence detection read-out system (Doctor Vida Pocket test) was successfully performed outside of the laboratory environment in less than 40 min using a proposed standard operation procedure. The results highlight that the use of the lyophilized LAMP reaction for food control purposes has the potential to meet the WHO-proposed REASSURED criteria.
Source: crossref · Quality: baseline
Research context
OpenAlex topics: Biosensors and Analytical Detection · Identification and Quantification in Food · Advanced biosensing and bioanalysis techniques
SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.
Affiliations
Hamburg School of Food Science, Institute of Food Chemistry, University of Hamburg, Grindelallee 117, 20146 Hamburg, Germany
Funding
Federal Ministry of Economic Affairs and Climate Action (BMWK)
References (41)
Show source-provided references
1. Omics approaches for food authentication10.1002/elps.201800004 ↗
2. Maquet, A., Lievens, A., Paracchini, V., Kaklamanos, G., de La Calle, B., Garlant, L., Papoci, S., Pietretti, D., Zdiniakova, T., and Breidbach, A. (2021). Results of an EU Wide Coordinated Control Plan to Establish the Prevalence of Fraudulent Practices in the Marketing of Herbs and Spices, Publications Office of the European Union. EUR30877EN; JRC126785.
3. Food and Agriculture Organization of the United Nations (FAO) (2024, September 28). The State of World Fisheries and Aquaculture 2022, Trade of Fisheries and Aquaculture Products. Available online: https://openknowledge.fao.org/server/api/core/bitstreams/9df19f53-b931-4d04-acd3-58a71c6b1a5b/content/sofia/2022/trade-of-aquatic-products.html.
4. Development of three real-time PCR assays to detect peanut allergen residue in processed food products10.1007/s00217-007-0797-3 ↗
5. REASSURED diagnostics to inform disease control strategies, strengthen health systems and improve patient outcomes10.1038/s41564-018-0295-3 ↗
6. LaBarre, P., Boyle, D., Hawkins, K., and Weigl, B. (2011, January 16). Instrument-free nucleic acid amplification assays for global health settings. In Proceedings of SPIE—The International Society for Optical Engineering, Orlando, FL, USA.10.1117/12.882868 ↗
7. Loop-mediated isothermal amplification of DNA10.1093/nar/28.12.e63 ↗
8. Loop-mediated isothermal amplification (LAMP): Principle, features, and future prospects10.1007/s12275-015-4656-9 ↗
9. Real-time kinetics and high-resolution melt curves in single-molecule digital LAMP to differentiate and study specific and non-specific amplification10.1093/nar/gkaa099 ↗
10. Herb and spice fraud; the drivers, challenges and detection10.1016/j.foodcont.2017.12.031 ↗
11. The application of loop-mediated isothermal amplification (LAMP) in food testing for bacterial pathogens and fungal contaminants10.1016/j.fm.2013.04.017 ↗
12. Detection of the food allergen celery via loop-mediated isothermal amplification technique10.1007/s00216-014-7873-x ↗
13. Cammilleri, G., Ferrantelli, V., Pulvirenti, A., Drago, C., Stampone, G., Del Rocio Quintero Macias, G., Drago, S., Arcoleo, G., Costa, A., and Geraci, F. (2020). Validation of a Commercial Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of Anisakis spp. DNA in Processed Fish Products. Foods, 9.10.3390/foods9010092 ↗
14. Njiru, Z.K. (2012). Loop-mediated isothermal amplification technology: Towards point of care diagnostics. PLoS Neglected Trop. Dis., 6.10.1371/journal.pntd.0001572 ↗
15. One simple DNA extraction device and its combination with modified visual loop-mediated isothermal amplification for rapid on-field detection of genetically modified organisms10.1021/ac301640p ↗
16. Rapid and low-cost identification of common sole (Solea solea) in the field using a fast DNA isolation protocol and loop-mediated isothermal amplification (LAMP)10.1016/j.jfca.2023.105166 ↗
17. Wan, J., Guo, J., Lu, Z., Bie, X., Lv, F., and Zhao, H. (2020). Development of a test kit for visual loop-mediated isothermal amplification of Salmonella in spiked ready-to-eat fruits and vegetables. J. Microbiol. Methods, 169.10.1016/j.mimet.2019.105830 ↗
18. Lyophilization and development of solid protein pharmaceuticals10.1016/S0378-5173(00)00423-3 ↗
19. Lyophilized visually readable loop-mediated isothermal reverse transcriptase nucleic acid amplification test for detection Ebola Zaire RNA10.1016/j.jviromet.2017.02.013 ↗
20. Characterization and Validation of a Lyophilized Loop-Mediated Isothermal Amplification Method for the Detection of Esox lucius10.1007/s12010-023-04799-x ↗
21. Colorimetric Detection of Horse Meat Based on Loop-Mediated Isothermal Amplification (LAMP)10.1007/s12161-019-01590-9 ↗
22. Rapid in-situ detection kit (RisK): Development of loop-mediated isothermal amplification (LAMP) assay for the rapid identification of selected invasive alien fish in Malaysian freshwaters10.1016/j.mcp.2020.101683 ↗
23. Establishment of a rapid method for skipjack tuna (Katsuwonus pelamis) authentication using molecular beacons in loop-mediated isothermal amplification10.1016/j.foodchem.2022.132365 ↗
24. An isothermal lab-on-phone test for easy molecular diagnosis of SARS-CoV-2 near patients and in less than 1 hour10.1016/j.ijid.2022.07.042 ↗
25. Papadakis, G., Pantazis, A.K., Fikas, N., Chatziioannidou, S., Tsiakalou, V., Michaelidou, K., Pogka, V., Megariti, M., Vardaki, M., and Giarentis, K. (2022). Portable real-time colorimetric LAMP-device for rapid quantitative detection of nucleic acids in crude samples. Sci. Rep., 12.10.1038/s41598-022-06632-7 ↗
26. (2024, September 30). Doctor Vida Pocket PCR Advertisement. Available online: https://www.stabvida.com/images/pricelists/doctor-vida-pocket-pcr.pdf?30012024.
27. (2010). Spices—Saffron (Crocus sativus L.)—Part 2: Test Methods. Standard No. ISO 3632-2:2010(en). Available online: https://www.dinmedia.de/de/norm/iso-3632-2/135451089.
28. (2009). Spices and Condiments—Determination of Total Ash. Standard No. ISO 928:2009(en). Available online: https://www.dinmedia.de/de/norm/une-iso-928/123990808.
29. (2019). Foodstuffs—DNA Barcoding of Fish and Fish Products Using Defined Mitochondrial Cytochrome b and Cytochrome c Oxidase I Gene Segments (Standard No. DIN CEN/TS 17303:2019-06).
30. Identification of the fish species of raw or cold-smoked salmon and salmon caviar by single-strand conformation polymorphism (SSCP) analysis10.1007/s00217-004-1067-2 ↗
31. Fast and User-Friendly Detection of Flatfish Species (Pleuronectes platessa and Solea solea) via Loop-Mediated Isothermal Amplification (LAMP)10.1021/acs.jafc.3c03917 ↗
32. Point-of-care suitable identification of the adulterants Carthamus tinctorius and Curcuma longa in Crocus sativus based on loop-mediated isothermal amplification (LAMP) and lateral-flow-assay (LFA)10.1016/j.foodcont.2023.109637 ↗
33. Factors affecting short-term and long-term stabilities of proteins10.1016/S0169-409X(00)00144-7 ↗
34. Cryoprotective Effect of Saccharides on Denaturation of Catalase by Freeze-Drying10.1248/cpb.39.1091 ↗
35. Optimization of Lyophilized LAMP and RT-PCR Reaction Mixes for Detection of Tuberculosis10.2478/ebtj-2020-0027 ↗
36. Rieder, J., Martin-Sanchez, P.M., Osman, O.A., Adrian-Kalchhauser, I., and Eiler, A. (2022). Detecting aquatic pathogens with field-compatible dried qPCR assays. J. Microbiol. Methods, 202.10.1016/j.mimet.2022.106594 ↗
37. Song, X., Coulter, F.J., Yang, M., Smith, J.L., Tafesse, F.G., Messer, W.B., and Reif, J.H. (2022). A lyophilized colorimetric RT-LAMP test kit for rapid, low-cost, at-home molecular testing of SARS-CoV-2 and other pathogens. Sci. Rep., 12.10.1038/s41598-022-11144-5 ↗
38. Prado, N.O., Marin, A.M., Lalli, L.A., Sanchuki, H.B.S., Wosniaki, D.K., Nardin, J.M., Morales, H.M.P., Blanes, L., Zanette, D.L., and Aoki, M.N. (2024). Development and evaluation of a lyophilization protocol for colorimetric RT-LAMP diagnostic assay for COVID-19. Sci. Rep., 14.10.1038/s41598-024-61163-7 ↗
39. A Genetic Lab-on-Phone Test for Point-of-Care Diagnostic of Lactose Intolerance near Patient and in less than 90 Minutes10.1093/jalm/jfad052 ↗
40. (2024, September 28). Visby Medical Sexual Health Test. Available online: https://www.visbymedical.com/sexual-health-test/.
41. Park, H.-J., Cho, H., Jung, H.S., Cho, B.H., and Lee, M.-Y. (2019). Development of a DNA isolation device using poly(3,4-dihydroxy-L-phenylalanine)-coated swab for on-site molecular diagnostics. Sci. Rep., 9.10.1038/s41598-019-44527-2 ↗