Loop-Mediated Isothermal Amplification for the Fast Detection of Bonamia ostreae and Bonamia exitiosa in Flat Oysters
Type | Article | ||||||||||||
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Date | 2024-02 | ||||||||||||
Language | English | ||||||||||||
Author(s) | Cano Irene1, Wood Gareth1, Stone David1, Noyer Mathilde2, Canier Lydie2, Arzul Isabelle2 | ||||||||||||
Affiliation(s) | 1 : The International Centre of Excellence for Aquatic Animal Health, Cefas Weymouth Laboratory, Weymouth, Dorset DT4 8UB, UK 2 : The Institut Français de Recherche pour l’Exploitation de la Mer Ifremer, RBE-SG2M-ASIM, Station de La Tremblade, Avenue de Mus de Loup, La Tremblade, 17390 Brest, France |
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Source | Pathogens (2076-0817) (MDPI AG), 2024-02 , Vol. 13 , N. 2 , P. 132 (15p.) | ||||||||||||
DOI | 10.3390/pathogens13020132 | ||||||||||||
Note | This article belongs to the Special Issue Aquatic Animal Pathogens: Epidemiology, Pathogenesis, Diagnosis, Preventive and Therapeutic Measures and Host–Pathogen Relationships | ||||||||||||
Keyword(s) | Bonamia ostreae, Bonamia exitiosa, bonamiosis, oyster, haplosporidia, LAMP, diagnostic, point-of-care test | ||||||||||||
Abstract | The haplosporidian parasites Bonamia ostreae (BO) and B. exitiosa (BE) are serious oyster pathogens. Two independent laboratories evaluated fluorescence real-time loop-mediated isothermal amplification (LAMP) assays for rapidly detecting these parasites. Specific LAMP assays were designed on the BO actin-1 and BE actin genes. A further generic assay was conceived on a conserved region of the 18S gene to detect both Bonamia species. The optimal reaction temperature varied from 65 to 67 °C depending on the test and instrument. Melting temperatures were 89.8–90.2 °C, 87.0–87.6 °C, and 86.2–86.6 °C for each of the BO, BE, and generic assays. The analytical sensitivity of these assays was 50 copies/µL in a 30 min run. The BO and BE test sensitivity was ~1 log lower than a real-time PCR, while the generic test sensitivity was similar to the real-time PCR. Both the BO and BE assays were shown to be specific; however, the generic assay potentially cross-reacts with Haplosporidium costale. The performance of the LAMP assays evaluated on samples of known status detected positives within 7–20 min with a test accuracy of 100% for the BO and generic tests and a 95.8% accuracy for BE. The ease of use, rapidity and affordability of these tests allow for field deployment. |
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