Detection of the infectious hematopoietic necrosis virus directly from infected fish tissues by dot blot hybridization with a non-radioactive probe.
A method was developed for the rapid diagnosis of the infectious hematopoietic necrosis virus (IHNV) based on dot blot hybridization with a non-radioactive probe. When the assay was developed through a color reaction both biotin- and alkaline phosphatase-labeled probes were highly specific for IHNV and the sensitivity reached to 20 pg of viral RNA. When the alkaline phosphatase-labeled probe was developed by a chemiluminiscent reaction, the sensitivity showed a five-fold increase to 4 pg of viral RNA. The procedures successfully detected IHNV directly from infected symptomatic and asymptomatic fishes exhibiting higher sensitivity than the traditional approaches involving viral propagation in cell cultures. Additional advantages of the method are its simplicity and it takes only about 6 h to carry out the procedures from the initial processing of the tissues to diagnosis.
González
M P
MP
Departamento de Ciencias Biologicas, Facultad de Quintica y Biologíu, Universidad de Santiago de Chile.
Sánchez
X
X
Ganga
M A
MA
López-Lastra
M
M
Jashés
M
M
Sandino
A M
AM
D000818:Animals; D002460:Cell Line; D003124:Colorimetry; D015342:DNA Probes / Q000737:chemistry / Q000378:metabolism; D008163:Luminescent Measurements; D009693:Nucleic Acid Hybridization; D017686:Oncorhynchus mykiss; D012367:RNA, Viral / Q000378:metabolism; D012209:Rhabdoviridae / Q000235:genetics* / Q000302:isolation & purification*; D018353:Rhabdoviridae Infections / Q000175:diagnosis* / Q000235:genetics* / Q000378:metabolism; D012680:Sensitivity and Specificity
D015342:DNA Probes; D012367:RNA, Viral