|
        |
     |
| |
J Vet Sci. 2006 Jun;7(2):167-176 |
|
| |
Characterization of Chinook head salmon embryo phenotypes of infectious salmon anemia virus by real-time RT-PCR
Khalid Munir* |
| |
Department of Pathology and Microbiology, Atlantic Veterinary College, University of Prince Edward Island, 550 University Avenue, Charlottetown, PEI C1A 4P3, Canada. kmunir@upei.ca |
| |
We have previously described the development of a onetube
SYBR Green real-time RT-PCR assay for the
detection and quantitation of infectious salmon anemia
virus (ISAV) in various biological samples. The twofold
aim of the present study was to verify that the optimized
SYBR Green real-time RT-PCR conditions could detect
ISAV isolates of different geographic origins, and to
analyze the growth patterns of the selected ISAV isolates
in the Chinook head salmon embryo (CHSE) -214 cells by
this assay to better characterize their CHSE-phenotypes.
A total of 24 ISAV isolates were used in this study. The
results indicated that the SYBR Green real-time RT-PCR
could detect ISAV of different geographic origins or
laboratory sources. The capacity of ISAV isolates to cause
cytopathic effects (CPE) in the CHSE-214 cell line, viral
titration of the infected CHSE-cell harvests, and analysis
of viral RNA levels in CHSE-214 cells at post-infection
day zero, 7 and 14 by SYBR Green real-time RT-PCR
confirmed the existence of three CHSE-phenotypes of
ISAV: replicating cytopathic, replicating non-cytopathic,
and non-replicating non-cytopathic. The identification of
these three CHSE- phenotypes of ISAV has important
implications from diagnostic and biological points of view.
|
|
| |
|