Abd-Eldaim Mohamed M, Wilkes Rebecca P, Thomas Kathy V, Kennedy Melissa A
Department of Comparative Medicine, University of Tennessee, Knoxville, 37996, USA.
Arch Virol. 2009;154(4):555-60. doi: 10.1007/s00705-009-0337-5. Epub 2009 Mar 1.
Feline calicivirus (FCV) is a common cause of upper respiratory tract disease in cats and is associated with interstitial pneumonia, oral ulceration and polyarthritis. Recently, outbreaks have involved a highly virulent FCV that leads to multisystemic signs. Virus isolation and conventional RT-PCR are the most common methods used for FCV diagnosis. However, real-time RT-PCR offers a rapid, sensitive, specific and easy tool for nucleic acid detection. The objective of this study was to design a TaqMan probe-based, real-time RT-PCR assay for detection of FCV. It was determined in our previous study that the first 120 nucleotides of the 5' region of the genome are highly conserved among FCV isolates. Primers and a probe specific for this region were designed for a real-time RT-PCR assay to detect FCV. Initial validation was done using 15 genetically diverse isolates. Also, 122 samples were tested by the new assay and virus isolation. The real-time RT-PCR assay was as sensitive and specific as virus isolation and was far more rapid. This real-time RT-PCR assay targeting the conserved 5' region of the genome is a fast, economical and accurate method for detection of FCV.
猫杯状病毒（FCV）是猫上呼吸道疾病的常见病因，与间质性肺炎、口腔溃疡和多关节炎有关。最近，疫情涉及一种高致病性FCV，可导致多系统症状。病毒分离和传统逆转录聚合酶链反应（RT-PCR）是用于FCV诊断的最常见方法。然而，实时RT-PCR为核酸检测提供了一种快速、灵敏、特异且简便的工具。本研究的目的是设计一种基于TaqMan探针的实时RT-PCR检测方法来检测FCV。我们之前的研究确定，基因组5'区域的前120个核苷酸在FCV分离株中高度保守。针对该区域设计了引物和探针，用于实时RT-PCR检测以检测FCV。最初使用15种遗传多样性不同的分离株进行了验证。此外，通过新方法和病毒分离对122份样本进行了检测。实时RT-PCR检测与病毒分离一样灵敏和特异，而且速度要快得多。这种针对基因组保守5'区域的实时RT-PCR检测方法是一种快速、经济且准确的FCV检测方法。
