Antemortem versus postmortem methods for detection of betanodavirus in Senegalese sole (Solea senegalensis).
The suitability of nested reverse transcription polymerase chain reaction (nRT-PCR) to detect betanodavirus in blood samples from naturally infected Senegalese sole (Solea senegalensis) was evaluated in comparison with other diagnostic methods. Results indicated that histologic examination of brain lesions could be regarded as the most consistent indicator of nodavirus infection in this species. The nRT-PCR showed low to moderate levels of detection; the best values were obtained in brain samples followed by blood samples. Inoculation of SSN-1 and SAF-1 cells with fish samples did not cause cytopathic effect, although virus was detected by reverse transcription polymerase chain reaction in approximately 25% of the SSN-1 inoculated wells. The efficiency of detection of the viral genome was dramatically increased by the use of nRT-PCR, reaching 90.6% of positives in brain samples and 84.4% in blood samples. The sensitivity and the negative predictive value of nRT-PCR in blood samples were slightly lower than those obtained using brain samples. Nevertheless, it is suggested that the advantage of being able to perform diagnosis on live fish adequately counterbalances the slightly lower sensitivity of nRT-PCR on blood samples. This technique is proposed as a useful tool, not only for the selection of nodavirus-free breeders but also to check the fish status during ongrowing.
Olveira
José Gabriel
JG
Unidad de Ictiopatología-Patalogía Viral, Instituto de Acuicultura, Dpto. Microbiología y Parasitología, Universidad de Santiago de Compostela, Santiago de Compostela, Spain.
Soares
Florbela
F
Engrola
Sofía
S
Dopazo
Carlos P
CP
Bandín
Isabel
I
D000818:Animals; D001927:Brain Diseases / Q000097:blood / Q000662:veterinary* / Q000821:virology; D002460:Cell Line; D003588:Cytopathogenic Effect, Viral; D005393:Fish Diseases / Q000097:blood / Q000821:virology*; D005412:Flatfishes; D030381:Nodaviridae / Q000235:genetics / Q000302:isolation & purification*; D011237:Predictive Value of Tests; D012327:RNA Virus Infections / Q000097:blood / Q000662:veterinary* / Q000821:virology; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity
D012367:RNA, Viral