Development of a quantitative polymerase chain reaction assay for detection of the aetiological agents of piscine lactococcosis.

Autor: Shahin K; Department of Medicine and Epidemiology, School of Veterinary Medicine, University of California, Davis, California, USA.; Aquatic Animal Diseases Laboratory, Aquaculture Department, National Institute of Oceanography and Fisheries, Suez, Egypt., Mukkatira K; California Department of Fish and Wildlife, Rancho Cordova, California, USA., Yazdi Z; Department of Medicine and Epidemiology, School of Veterinary Medicine, University of California, Davis, California, USA., Richey C; California Department of Fish and Wildlife, Rancho Cordova, California, USA., Kwak K; California Department of Fish and Wildlife, Rancho Cordova, California, USA., Heckman TI; Department of Medicine and Epidemiology, School of Veterinary Medicine, University of California, Davis, California, USA., Mohammed HH; Department of Aquatic Animal Medicine and Management, Faculty of Veterinary Medicine, Assiut University, Assiut, Egypt., Ortega C; Centro de Investigación y Estudios Avanzados en Salud Animal (CIESA), Facultad de Medicina Veterinaria y Zootecnia (FMVZ), Universidad Autónoma del Estado de México (UAEM), Toluca, Mexico., Avendaño-Herrera R; Laboratorio de Patología de Organismos Acuáticos y Biotecnología Acuícola, Universidad Andrés Bello and Centro FONDAP INCAR, Viña del Mar, Chile., Keleher B; Kennebec River Biosciences, Richmond, Maine, USA., Hyatt MW; Wildlife Conservation Society, New York Aquarium, Brooklyn, New York, USA., Drennan JD; Colorado Parks and Wildlife, Brush, Colorado, USA., Adkison M; California Department of Fish and Wildlife, Rancho Cordova, California, USA., Griffin MJ; Department of Pathobiology and Population Medicine, College of Veterinary Medicine, Mississippi State University, Stoneville, Mississippi, USA., Soto E; Department of Medicine and Epidemiology, School of Veterinary Medicine, University of California, Davis, California, USA.
Jazyk: angličtina
Zdroj: Journal of fish diseases [J Fish Dis] 2022 Jun; Vol. 45 (6), pp. 847-859. Date of Electronic Publication: 2022 Mar 20.
DOI: 10.1111/jfd.13610
Abstrakt: Piscine lactococcosis is an emergent bacterial disease that is associated with high economic losses in many farmed and wild aquatic species worldwide. Early and accurate detection of the causative agent of piscine lactococcosis is essential for management of the disease in fish farms. In this study, a TaqMan quantitative polymerase chain reaction (qPCR) targeting the 16S-23S rRNA internal transcribed spacer region was developed and validated. Validation of the qPCR was performed with DNA of previously typed L. petauri and L. garvieae recovered from different aquatic hosts from distinct geographical locations, closely related bacterial species and common pathogens in trout aquaculture. Further diagnostic sensitivity and specificity was investigated by screening of fish, water and faecal samples. The developed qPCR assay showed high specificity, sensitivity and accuracy in detection of L. petauri and L. garvieae with lack of signals from non-target pathogens, and in screening of rainbow trout (Oncorhynchus mykiss) posterior kidney and environmental samples. The detection limit of the qPCR was four amplicon copies. Moreover, the sensitivity of the qPCR assay was not affected by presence of non-target DNA from either fish or environmental samples. The robustness, specificity and sensitivity of the developed qPCR will facilitate fast and accurate diagnosis of piscine lactococcosis to establish appropriate control measures in fish farms and aquaria.
(© 2022 John Wiley & Sons Ltd.)
Databáze: MEDLINE
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