Abstract
Atlantic salmon (Salmo salar) is consumed worldwide for its flavor and high nutritional value. However, because S. salar is difficult and costly to breed and has low yield, it remains difficult to meet the global demand. Rainbow trout (Oncorhynchus mykiss) is closely related to S. salar, and their flesh is similar in appearance. Because the price difference between these two fish species is large, O. mykiss is sometimes sold as S. salar to maximize profits, which is a violation of consumer rights and could pose health risks. Freshwater farmed O. mykiss is at higher risk of infection by parasites that can easily infect humans, whereas those carried by marine-cultured S. salar cannot. Thus, there is an urgent need for rapid, simple, and sensitive detection methods to differentiate these two species. In this study, we developed a method that combines isothermal recombinase polymerase amplification and lateral flow strip (RPA-LFS) to identify S. salar. A standard sample of S. salar was used as a template, and the myoglobin gene was selected as a target gene to establish the RPA-LFS detection method. Frozen fish samples were purchased online and analyzed using the RPA-LFS system; the results were compared with those obtained using PCR amplification and cytochrome oxidase I (COI) gene sequencing. The RPA-LFS system successfully performed amplification at 37 °C, and the entire detection process required approximately 20 min. The minimum detection limit was 102 ng/mL or 103 copies/mL. The detection results for clinical samples were consistent with those obtained using DNA barcoding technology (32/36). Together, these findings indicate that the detection system developed in this study is applicable for rapid, high-precision, sensitive identification of S. salar.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Explore related subjects
Discover the latest articles and news from researchers in related subjects, suggested using machine learning.Availability of Data and Materials
Not applicable.
Code Availability
Not applicable.
References
Abdullah A, Rehbein H (2016) The differentiation of tuna (family: Scombridae) products through the PCR-based analysis of the cytochrome b gene and parvalbumin introns. J Sci Food Agric 96(2):456–464. https://doi.org/10.1002/jsfa.7111
Aidos I, Lie O, Espe M (1999) Collagen content in farmed Atlantic salmon (Salmo salar L.). J Agric Food Chem 47(4):1440–1444. https://doi.org/10.1021/jf980761
Carrera E, García T, Céspedes A, González I, Fernández A, Asensio LM, Hernández PE, Martín R (2000) Identification of smoked Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss) using PCR-restriction fragment length polymorphism of the p53 gene. J AOAC Int 83(2):341–346
Carrera E, García T, Céspedes A, González I, Sanz B, Hernández PE, Martín R (1998) Identification of Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss) by using polymerase chain reaction amplification and restriction analysis of the mitochondrial cytochrome b gene. J Food Prot 61(4):482–486. https://doi.org/10.4315/0362-028x-61.4.482
Carrera E, Martín R, García T, González I, Sanz B, Hernández PE (1996) Development of an enzyme-linked immunosorbent assay for the identification of smoked salmon (Salmo salar), trout (Oncorhynchus mykiss) and bream (Brama raii). J Food Prot 59(5):521–524. https://doi.org/10.4315/0362-028x-59.5.521
Chen Z, Wu T, Xiang C, Xu X, Tian X (2019) Rapid identification of rainbow trout adulteration in Atlantic salmon by Raman spectroscopy combined with machine learning. Molecules 24(15). https://doi.org/10.3390/molecules24152851
Daher RK, Stewart G, Boissinot M, Bergeron MG (2016) Recombinase polymerase amplification for diagnostic applications. Clin Chem 62(7):947–958. https://doi.org/10.1373/clinchem.2015.245829
Dai T, Yang X, Hu T, Jiao B, Xu Y, Zheng X, Shen D (2019) Comparative evaluation of a novel recombinase polymerase amplification-lateral flow dipstick (RPA-LFD) assay, LAMP, conventional PCR, and leaf-disc baiting methods for detection of Phytophthora sojae. Front Microbiol 10:1884. https://doi.org/10.3389/fmicb.2019.01884
Dalvin S, Glover KA, Sørvik AG, Seliussen BB, Taggart JB (2010) Forensic identification of severely degraded Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss) tissues. Investig Genet 1(1):12. https://doi.org/10.1186/2041-2223-1-12
Espiñeira M, González-Lavín N, Vieites JM, Santaclara FJ (2008) Authentication of anglerfish species (Lophius spp) by means of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and forensically informative nucleotide sequencing (FINS) methodologies. J Agric Food Chem 56(22):10594–10599. https://doi.org/10.1021/jf801728q
Feng J, Wu Z, Xie X, Dai Z, Liu S (2017) A real-time polymerase chain reaction method for the identification of four commercially important salmon and trout species. Mitochondrial DNA A DNA Mapp Seq Anal 28(1):104–111. https://doi.org/10.3109/19401736.2015.1111346
Gu S, Deng X, Shi Y, Cai Y, Huo Y, Guo D, Han F (2020) Identification of peptide biomarkers for authentication of Atlantic salmon and rainbow trout with untargeted and targeted proteomics approaches and quantitative detection of adulteration. J Chromatogr B Analyt Technol Biomed Life Sci 1155:122194. https://doi.org/10.1016/j.jchromb.2020.122194
Hjorthaug HS, Gervin K, Mowinckel P, Munthe-Kaas MC (2018) Exploring the influence from whole blood DNA extraction methods on Infinium 450K DNA methylation. PLoS ONE 13(12):e0208699. https://doi.org/10.1371/journal.pone.0208699
Huang, M., Xiong, X., Xu, WJ., Li, Y., Cao, M., Xiong, XH. (2020). Development and application of a dual PCR method for detection of Atlantic salmon and rainbow trout. Modern Food Science and Technology, V.36;No.256(12), 280–288
Levipan HA, Avendaño-Herrera R (2021) Assessing the impacts of skin mucus from Salmo salar and Oncorhynchus mykiss on the growth and in vitro infectivity of the fish pathogen Piscirickettsia salmonis. J Fish Dis 44(2):181–190. https://doi.org/10.1111/jfd.13275
Liu L, Li R, Zhang R, Wang J, An Q, Han Q, Wang J, Yuan W (2019) Rapid and sensitive detection of Mycoplasma hyopneumoniae by recombinase polymerase amplification assay. J Microbiol Methods 159:56–61. https://doi.org/10.1016/j.mimet.2019.02.015
Lock EJ, Ornsrud R, Aksnes L, Spanings FA, Waagbø R, Flik G (2007) The vitamin D receptor and its ligand 1alpha,25-dihydroxyvitamin D3 in Atlantic salmon (Salmo salar). J Endocrinol 193(3):459–471. https://doi.org/10.1677/joe-06-0198
Ma B, Li J, Chen K, Yu X, Sun C, Zhang M (2020) Multiplex recombinase polymerase amplification assay for the simultaneous detection of three foodborne pathogens in seafood. Foods 9(3). https://doi.org/10.3390/foods9030278
MacWilliams C, Johnson G, Groman D, Kibenge FS (2007) Morphologic description of infectious salmon anaemia virus (ISAV)-induced lesions in rainbow trout Oncorhynchus mykiss compared to Atlantic salmon Salmo salar. Dis Aquat Organ 78(1):1–12. https://doi.org/10.3354/dao01866
Marchetti P, Mottola A, Piredda R, Ciccarese G, Di Pinto A (2020) Determining the authenticity of shark meat products by DNA sequencing. Foods 9 (9). 10.3390/foods9091194
Pappalardo AM, Copat C, Raffa A, Rossitto L, Grasso A, Fiore M, Ferrante M, Ferrito V (2020) Fish-based baby food concern-from species authentication to exposure risk assessment. Molecules 25(17). https://doi.org/10.3390/molecules25173961
Peng J, Larondelle Y, Pham D, Ackman RG, Rollin X (2003) Polyunsaturated fatty acid profiles of whole body phospholipids and triacylglycerols in anadromous and landlocked Atlantic salmon (Salmo salar L.) fry. Comp Biochem Physiol B Biochem Mol Biol 134(2):335–348. https://doi.org/10.1016/s1096-4959(02)00263-4
Piepenburg O, Williams CH, Stemple DL, Armes NA (2006) DNA detection using recombination proteins. PLoS Biol 4(7):e204. https://doi.org/10.1371/journal.pbio.0040204
Rasmussen Hellberg RS, Naaum AM, Handy SM, Hanner RH, Deeds JR, Yancy HF, Morrissey MT (2011) Interlaboratory evaluation of a real-time multiplex polymerase chain reaction method for identification of salmon and trout species in commercial products. J Agric Food Chem 59(3):876–884. https://doi.org/10.1021/jf103241y
Shokralla S, Hellberg RS, Handy SM, King I, Hajibabaei M (2015) A DNA mini-barcoding system for authentication of processed fish products. Sci Rep 5:15894. https://doi.org/10.1038/srep15894
Shulman BS, Ieshko EP, Shurov IL, Ensen BO, Ensen A (2004) Parasite fauna of the landlocked Atlantic salmon Salmo salar L. in the basin of Namsen river. Parazitologiia 38(1):94–97
Wang L, Zhao P, Si X, Li J, Dai X, Zhang K, Gao S, Dong J (2019) Rapid and specific detection of Listeria monocytogenes with an isothermal amplification and lateral flow strip combined method that eliminates false-positive signals from primer-dimers. Front Microbiol 10:2959. https://doi.org/10.3389/fmicb.2019.02959
Wang Y, Jiao WW, Wang Y, Wang YC, Shen C, Qi H, Shen AD (2020) Graphene oxide and self-avoiding molecular recognition systems-assisted recombinase polymerase amplification coupled with lateral flow bioassay for nucleic acid detection. Mikrochim Acta 187(12):667. https://doi.org/10.1007/s00604-020-04637-5
Ward LN, Bej AK (2006) Detection of Vibrio parahaemolyticus in shellfish by use of multiplexed real-time PCR with TaqMan fluorescent probes. Appl Environ Microbiol 72(3):2031–2042. https://doi.org/10.1128/aem.72.3.2031-2042.2006
Yang Y, Qin X, Song Y, Zhang W, Hu G, Dou Y, Li Y, Zhang Z (2017) Development of real-time and lateral flow strip reverse transcription recombinase polymerase amplification assays for rapid detection of peste des petits ruminants virus. Virol J 14(1):24. https://doi.org/10.1186/s12985-017-0688-6
Yuan QB, Huang YM, Wu WB, Zuo P, Hu N, Zhou YZ, Alvarez PJJ (2019) Redistribution of intracellular and extracellular free & adsorbed antibiotic resistance genes through a wastewater treatment plant by an enhanced extracellular DNA extraction method with magnetic beads. Environ Int 131:104986. https://doi.org/10.1016/j.envint.2019.104986
Acknowledgements
The authors acknowledge the support of the Jiangsu Key Laboratory of Marine Pharmaceutical Compound Screening for the provision of standard material samples.
Funding
This study was funded by the Sericulture Industry Technology Research Institute, Agriculture Research System (CARS-18-ZJ0207), Zhenjiang Agricultural Technology (NY2018013), and Jiangsu University of Science and Technology (1732931603).
Author information
Authors and Affiliations
Contributions
Weiguo Zhao and Lei Wang designed the study. Lei Wang, Danyan Zheng, Huahua Wu, Shishi Qi, Xiangdong Xin, and Peng Guo conducted the research. Lei Wang, Michael Ackah, and Weiguo Zhao wrote the manuscript. Weiguo Zhao directed the project.
Corresponding author
Ethics declarations
Ethics Approval
All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.
Informed Consent
Not applicable.
Consent to Participate
Not applicable.
Consent for Publication
Not applicable.
Conflict of Interest
Lei Wang declares no conflict of interest. Danyan Zheng declares no conflict of interest. Michael Ackah declares no conflict of interest. Huahua Wu declares no conflict of interest. Shishi Qi declares no conflict of interest. Xiangdong Xin declares no conflict of interest. Peng Guo declares no conflict of interest. Weiguo Zhao declares no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Lei Wang, Danyan Zheng, and Michael Ackah contributed equally to this work.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Wang, L., Zheng, D., Ackah, M. et al. Rapid Identification of Salmo salar Using a Combined Isothermal Recombinase Polymerase Amplification–Lateral Flow Strip Approach. Food Anal. Methods 15, 401–412 (2022). https://doi.org/10.1007/s12161-021-02128-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12161-021-02128-8