Species-specific loop-mediated isothermal amplification (LAMP) assays for the detection of three blood flukes affecting bluefin tuna aquaculture
4 affiliations
- 1Department of Biology, School of Science, RMIT University, Melbourne, VIC 3083 Australia
- 2Aquaculture Research Institute, Kindai University, Wakayama, 649-2211 Japan
- 3Department of Surgery, Austin Health, The University of Melbourne, Heidelberg, VIC 3084 Australia
- 4Institute for Marine and Antarctic Sciences (IMAS), University of Tasmania, Locked Bag 1370, Launceston, TAS 7250 Australia
Abstract
Blood flukes of the genus Cardicola (Digenea: Aporocotylidae) are well-documented health threats to farmed and ranched bluefin tuna (BFT), where infections can result in substantial economic losses. While recombinase polymerase assays (RPA) have been developed for two species of Cardicola, the high degree of sequence conservation across all three species reported from BFT requires an alternative method to enable confident species-level discrimination. The aims of this study were to design, optimise and validate loop-mediated isothermal amplification (LAMP) for the detection of the three Cardicola species which infect BFT, and apply the assays to Pacific bluefin tuna (PBT) cultured in Japan. Species-specific LAMP assays were successfully designed, optimised and validated for C. forsteri, C. opisthorchis and C. orientalis targeting ITS-2 ribosomal DNA. In addition, these assays were applied to DNA from three sample types targeting sites of significant pathology, the heart, gill necropsy and gill biopsy of 15 PBT. Results were compared to outputs of quantitative polymerase chain reactions (qPCR), the gold standard for blood fluke detection in ranched Australian SBT, where LAMP assays were shown to be more sensitive. This research constitutes the first development and application of species-specific LAMP diagnostics for the three described Cardicola species infecting BFT, providing a rapid, cost-effective and accessible alternative to standard molecular diagnostics, informing management and treatment decisions.
Supplementary Information: The online version contains supplementary material available at https://doi.org/10.1007/s00436-026-08775-2.
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BibTeX
@article{carabott2026speciesspecific,
title = {{Species-specific loop-mediated isothermal amplification (LAMP) assays for the detection of three blood flukes affecting bluefin tuna aquaculture}},
author = {Melissa J. Carabott and Cecilia Power and Sho Shirakashi and Paul A. Ramsland and Barbara F. Nowak and Nathan J. Bott},
journal = {Parasitology Research},
year = {2026},
month = sep,
doi = {10.1007/s00436-026-08775-2},
url = {https://doi.org/10.1007/s00436-026-08775-2},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Species-specific loop-mediated isothermal amplification (LAMP) assays for the detection of three blood flukes affecting bluefin tuna aquaculture
AU - Melissa J. Carabott
AU - Cecilia Power
AU - Sho Shirakashi
AU - Paul A. Ramsland
AU - Barbara F. Nowak
AU - Nathan J. Bott
PY - 2026
DA - 2026/09/30
JO - Parasitology Research
DO - 10.1007/s00436-026-08775-2
UR - https://doi.org/10.1007/s00436-026-08775-2
AB - Blood flukes of the genus Cardicola (Digenea: Aporocotylidae) are well-documented health threats to farmed and ranched bluefin tuna (BFT), where infections can result in substantial economic losses. While recombinase polymerase assays (RPA) have been developed for two species of Cardicola, the high degree of sequence conservation across all three species reported from BFT requires an alternative method to enable confident species-level discrimination. The aims of this study were to design, optimise and validate loop-mediated isothermal amplification (LAMP) for the detection of the three Cardicola species which infect BFT, and apply the assays to Pacific bluefin tuna (PBT) cultured in Japan. Species-specific LAMP assays were successfully designed, optimised and validated for C. forsteri, C. opisthorchis and C. orientalis targeting ITS-2 ribosomal DNA. In addition, these assays were applied to DNA from three sample types targeting sites of significant pathology, the heart, gill necropsy and gill biopsy of 15 PBT. Results were compared to outputs of quantitative polymerase chain reactions (qPCR), the gold standard for blood fluke detection in ranched Australian SBT, where LAMP assays were shown to be more sensitive. This research constitutes the first development and application of species-specific LAMP diagnostics for the three described Cardicola species infecting BFT, providing a rapid, cost-effective and accessible alternative to standard molecular diagnostics, informing management and treatment decisions. Supplementary Information: The online version contains supplementary material available at https://doi.org/10.1007/s00436-026-08775-2.
SN - 0932-0113
SN - 1432-1955
ER -