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Multiplex PCR assay for the accurate and rapid detection and differentiation of Lactococcus garvieae and L. petauri.
Ustaoglu, Dilek; Öztürk, Rafet Çagri; Ture, Mustafa; Colussi, Silvia; Pastorino, Paolo; Vela, Ana Isabel; Kotzamanidis, Charalampos; Volpatti, Donatella; Acutis, Pier Luigi; Altinok, Ilhan.
Afiliación
  • Ustaoglu D; Department of Fisheries Technology Engineering, Faculty of Marine Sciences, Karadeniz Technical University, Trabzon, Turkey.
  • Öztürk RÇ; Aquatic Animal Health and Molecular Genetic Lab, Karadeniz Technical University, Trabzon, Turkey.
  • Ture M; Department of Fisheries Technology Engineering, Faculty of Marine Sciences, Karadeniz Technical University, Trabzon, Turkey.
  • Colussi S; Aquatic Animal Health and Molecular Genetic Lab, Karadeniz Technical University, Trabzon, Turkey.
  • Pastorino P; Department of Fish Health, Central Fisheries Research Institute, Trabzon, Turkey.
  • Vela AI; Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle d'Aosta, Turin, Italy.
  • Kotzamanidis C; Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle d'Aosta, Turin, Italy.
  • Volpatti D; VISAVET and Department of Animal Health, Universidad Complutense de Madrid, Madrid, Spain.
  • Acutis PL; Veterinary Research Institute, ELGO-DIMITRA, Thessaloniki, Greece.
  • Altinok I; Department of Agricultural, Food, Environmental and Animal Sciences (DI4A), University of Udine, Udine, Italy.
J Fish Dis ; : e14004, 2024 Aug 04.
Article en En | MEDLINE | ID: mdl-39097825
ABSTRACT
Lactococcosis is a common bacterial fish disease caused by Lactococcus garvieae, L. petauri and L. formosensis. Although there are different PCR-based techniques to identify the etiological agent, none of these can differentiate these two bacteria without sequencing PCR-amplified fragments. In the present study, we developed a multiplex PCR assay for simultaneous detection and differentiation of L. garvieae and L. petauri. The specificity of the primers was validated against the bacterial DNA of the targeted and non-targeted bacteria. The sizes of the PCR amplicons were obtained as 204 bp for the DUF1430 domain-containing protein gene of L. garvieae, 465 bp for the Lichenan permease IIC component gene of L. petauri, and 302 bp for the teichoic acid biosynthesis protein F gene of both L. garvieae and L. petauri. The PCR amplicons were clearly separated by agarose gel electrophoresis. The multiplex PCR assay did not produce any amplification products with the DNA of the non-targeted bacteria. The multiplex PCR detection limits for L. garvieae and L. petauri were 5 and 4 CFU in pure culture and 50 and 40 CFU/g in spiked tissue samples, respectively. It takes less than 2 h from plate-cultured bacteria and 3 h from tissue samples to get results. In conclusion, the developed multiplex PCR assay is a rapid, specific, accurate, and cost-effective method for the detection and differentiation of L. garvieae and L. petauri and is suitable to be used for routine laboratory diagnosis of L. garvieae and L. petauri.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: J Fish Dis Asunto de la revista: BIOLOGIA / MEDICINA VETERINARIA Año: 2024 Tipo del documento: Article País de afiliación: Turquía Pais de publicación: Reino Unido
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: J Fish Dis Asunto de la revista: BIOLOGIA / MEDICINA VETERINARIA Año: 2024 Tipo del documento: Article País de afiliación: Turquía Pais de publicación: Reino Unido