Development of loop-mediated isothermal amplification (LAMP) assay for rapid and direct screening of yellowfin tuna (Thunnus albacares) in commercial fish products

Abstract

Tuna is one of the most widely consumed fish on the European market, being available in various consumable options. Among them, Thunnus albacares, also called yellowfin tuna, is a delicacy and is consumed by millions of people around the world. Due to its comparatively high cost and demand, it is more vulnerable to fraud, where low-cost tuna or other fish varieties might be replaced for economic gain. In this study, a loop-mediated isothermal amplification (LAMP) assay was developed and validated for targeting the mitochondrial cytochrome b gene for fast and direct detection of Thunnus albacares, which is a valuable tuna species. The analytical specificity was confirmed using 18 target samples (Thunnus albacares) and 18 samples of non-target fish species. The analytical sensitivity of the LAMP assay was 540 fg DNA per reaction. In addition, a simple and direct swab method without time-consuming nucleic acid extraction procedures and the necessity for cost-intensive laboratory equipment was performed that allowed LAMP detection of Thunnus albacares samples within 13 minutes. Due to its high specificity and sensitivity, the LAMP assay can be used as a rapid and on-site screening method for identifying Thunnus albacares, potentially providing a valuable monitoring tool for food authenticity control by the authorities.

Parvalbumin Gene: A Valuable Marker for Pike Authentication and Allergen Risk Assessment

“…Furthermore, another reaction mixture containing a polymerase with higher activity or providing better reaction conditions for the polymerase could be used. Ali et al have achieved the detection of Thunnus albacares in less than 15 min when using the GspSSD Isothermal Mastermix (ISO-001) (OptiGene Ltd., UK). The same Mastermix was also used by Abdulmawjood et al who obtained the results of the LAMP assay in 30 min.…”

“…While the PCR method has been used for fish identification for a long time, the LAMP method is still used more for the detection of fish parasites and other microorganisms. 49−52 To the best of our knowledge, the use of LAMP for the identification of the fish itself has been published only for the members of salmon, 53,54 trout, 55 tuna, 29,56 eel, 57 cod, 58 common sole, 59 Arothorn or Diodon genus. 60,61 Thus, we believe that the present paper is the first to propose a LAMP protocol for pike species identification and, moreover, one of the few that use a nuclear gene as a marker since most published protocols have focused on mitochondrial cytochrome b.…”

Rapid analysis technologies with chemometrics for food authenticity field: A review

“…LAMP-based fish species-specific identification studies serve two purposes. One is to establish a specific analysis method based on LAMP for high-value species, such as Skipjack tuna, Thunnus albacares , and Salmo salar , to identify adulteration in the corresponding processed products ( Ali et al, 2022 ; Li et al, 2022 ; Xiong et al, 2021 ). Another is the use of LAMP to quickly identify toxic fish species (diodon, arothron and takifugu) in commercially available products, effectively reducing the incidence of poisonings ( Xie et al, 2022a , 2022b , 2022c ).…”

Fast and User-Friendly Detection of Flatfish Species (Pleuronectes platessa and Solea solea) via Loop-Mediated Isothermal Amplification (LAMP)

“…Thus, the developed LAMP assays are much more suitable for routine analysis in the laboratory, for in-field testing, and for limited-resource settings than conventional PCR-based methods. LAMP has been successfully developed on cytb sequences for a wide range of animal-derived foods, including fish species such as Salmo salar and Oncorhynchus mykiss , belonging to the genus Gadus , ,, Katsuwonus pelamis, , Thunnus albacares, and Anguilla anguilla …”

“…16 Thus, the developed LAMP assays are much more suitable for routine analysis in the laboratory, for in-field testing, and for limited-resource settings than conventional PCR-based methods. LAMP has been successfully developed on cytb sequences for a wide range of animal-derived foods, including fish species such as Salmo salar and Oncorhynchus mykiss 15,16 belonging to the genus Gadus, 10,17,18 Katsuwonus pelamis, 19,20 Thunnus albacares, 21 and Anguilla anguilla. 22 In this study, we developed a loop-mediated isothermal amplification assay for rapid species detection of Pleuronectes platessa and Solea solea, economically important fish species in Europe belonging to the order of flatfish (Pleuronectiformes).…”