Munir Khalid
Department of Pathology and Microbiology, Atlantic Veterinary College, University of Prince Edward Island, 550 University Avenue, Charlottetown, PEI C1A 4P3, Canada.
J Vet Sci. 2006 Jun;7(2):167-76. doi: 10.4142/jvs.2006.7.2.167.
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.
我们之前描述了一种单管SYBR Green实时逆转录聚合酶链反应（RT-PCR）检测方法的开发，用于检测和定量各种生物样品中的传染性鲑鱼贫血病毒（ISAV）。本研究的双重目的是验证优化后的SYBR Green实时RT-PCR条件能否检测不同地理来源的ISAV分离株，并通过该检测方法分析所选ISAV分离株在奇努克鲑鱼头肾胚胎（CHSE）-214细胞中的生长模式，以更好地表征它们的CHSE表型。本研究共使用了24株ISAV分离株。结果表明，SYBR Green实时RT-PCR能够检测不同地理来源或实验室来源的ISAV。通过SYBR Green实时RT-PCR对ISAV分离株在CHSE-214细胞系中引起细胞病变效应（CPE）的能力、感染的CHSE细胞收获物的病毒滴定以及感染后第0天、7天和14天CHSE-214细胞中病毒RNA水平的分析，证实了ISAV存在三种CHSE表型：复制性细胞病变型、复制性非细胞病变型和非复制性非细胞病变型。从诊断和生物学角度来看，鉴定出ISAV的这三种CHSE表型具有重要意义。
