Development and validation of a short-time cell culture and multiplex reverse transcriptase polymerase chain reaction assay for infectious pancreatic necrosis virus in Mexican farm-sampled rainbow trout.
J Aquat Anim Health
; 21(3): 167-72, 2009 Sep.
Article
en En
| MEDLINE
| ID: mdl-20043402
ABSTRACT
The infectious pancreatic necrosis virus (IPNV) affects several species of freshwater and marine fish. In Mexico, IPNV has an important impact on farming of rainbow trout Oncorhynchus mykiss; however, IPNV distribution in Mexico is unclear. The diagnosis of IPNV is laborious; usually it is based on isolation tests in cell culture followed by immunological identification using techniques of serum neutralization, immunofluorescence, or enzyme-linked immunosorbent assay. It has recently been demonstrated that reverse transcriptase polymerase chain reaction (RT-PCR) is an adequate method for the detection of aquatic birnaviruses. However, its diagnostic use is still limited because very low titers of viable virus cannot be easily detected. In this study, a combination of short-time cell culture and multiplex RT-PCR was established for the diagnosis of IPNV in rainbow trout obtained from farms in the state of Mexico. Three primer sets were used in a single reaction in the multiplex RT-PCR to increase the probability of identifying all serotypes of IPNV serogroup A as well as to help prevent a false-negative result. This approach was able to identify samples with an IPNV concentration of just 0.01 tissue culture infective dose with 50% endpoint (TCID50)/mL, and it identified more infected fish than RT-PCR alone or first-passage cell culture alone. Moreover, this technique made the same identifications as second-passage cell culture but in approximately 30% of the time needed for second-passage cell culture. Consequently, the time and cost efficiency of IPNV diagnosis were greatly reduced.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Oncorhynchus mykiss
/
Virus de la Necrosis Pancreática Infecciosa
/
Infecciones por Birnaviridae
/
Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
/
Enfermedades de los Peces
Tipo de estudio:
Diagnostic_studies
/
Prognostic_studies
Límite:
Animals
País/Región como asunto:
Mexico
Idioma:
En
Revista:
J Aquat Anim Health
Asunto de la revista:
MEDICINA VETERINARIA
Año:
2009
Tipo del documento:
Article