A combined RT‐PCR and dot‐blot hybridization method reveals the coexistence of SJNNV and RGNNV betanodavirus genotypes in wild meagre (Argyrosomus regius) |
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Authors: | B Lopez‐Jimena N Cherif E Garcia‐Rosado C Infante I Cano D Castro S Hammami JJ Borrego MC Alonso |
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Institution: | 1. IFAPA Centro El Toru?o, Junta de Andalucía. El Puerto de Santa María, Cádiz, Spain;2. Institut de la Recherche Veterinaire de Tunisie, Rue de Djebel Lakhdhar, La Rabta Tunis, Tunisie;3. Departamento de Microbiología, Facultad de Ciencias, Universidad de Málaga, Málaga, Spain |
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Abstract: | Aims: To detect the possible coexistence of striped jack nervous necrosis virus (SJNNV) and red‐spotted grouper nervous necrosis virus (RGNNV) genotypes in a single fish, a methodology based on the combination of PCR amplification and blot hybridization has been developed and applied in this study. Methods and Results: The degenerate primers designed for the PCR procedure target the T4 region within the capsid gene, resulting in the amplification of both genotypes. The subsequent hybridization of these amplification products with two different specific digoxigenin‐labelled probes resulted in the identification of both genotypes separately. The application of the RT‐PCR protocol to analyse blood samples from asymptomatic wild meagre (Argyrosomus regius) specimens has shown a 46·87% of viral nervous necrosis virus carriers. The combination of RT‐PCR and blot hybridization increases the detection rate up to 90·62%, and, in addition, it has shown the coexistence of both genotypes in 18 out of the 32 specimens analysed (56·25%). Conclusions: This study reports the coexistence of betanodaviruses belonging to two different genotypes (SJNNV and RGNNV) in wild fish specimens. Significance and Impact of the Study: This is the first report demonstrating the presence of SJNNV and RGNNV genotypes in the same specimen. This study also demonstrates a carrier state in this fish species for the first time. |
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Keywords: | betanodavirus degenerate primers dot‐blot hybridization RGNNV RT‐PCR SJNNV |
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