Simons Fermin A, Vennema Harry, Rofina Jaime E, Pol Jan M, Horzinek Marian C, Rottier Peter J M, Egberink Herman F
Institute of Virology, Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3584 CL Utrecht, The Netherlands.
J Virol Methods. 2005 Mar;124(1-2):111-6. doi: 10.1016/j.jviromet.2004.11.012. Epub 2004 Dec 21.
A reverse transcriptase polymerase chain reaction (RT-PCR) for the detection of feline coronavirus (FCoV) messenger RNA in peripheral blood mononuclear cells (PBMCs) is described. The assay is evaluated as a diagnostic test for feline infectious peritonitis (FIP). It is based on a well-documented key event in the development of FIP: the replication of virulent FCoV mutants in monocytes/macrophages. To detect most feline coronavirus field strains, the test was designed to amplify subgenomic mRNA of the highly conserved M gene. The test was applied to 1075 feline blood samples (424 from healthy, 651 from sick cats suspected of FIP) and returned 46% of the diseased cats as positive for feline coronavirus mRNA in their peripheral blood cells; of the healthy cats, 5% tested positive. Of a group of 81 animals in which FIP had been confirmed by post-mortem examination, 75 (93%) tested positive, whereas 17 cats with different pathologies (non-FIP cases) all tested negative. In view of the low rate of false-positive results (high specificity) the mRNA RT-PCR may be a valuable addition to the diagnostic arsenal for FIP.
本文描述了一种用于检测外周血单个核细胞(PBMC)中猫冠状病毒(FCoV)信使核糖核酸(mRNA)的逆转录聚合酶链反应(RT-PCR)。该检测方法被评估为猫传染性腹膜炎(FIP)的诊断试验。它基于FIP发展过程中一个有充分文献记载的关键事件:毒性FCoV突变体在单核细胞/巨噬细胞中的复制。为了检测大多数猫冠状病毒野毒株,该检测方法设计用于扩增高度保守的M基因的亚基因组mRNA。该检测方法应用于1075份猫血样(424份来自健康猫,651份来自疑似FIP的病猫),结果显示,46%的病猫外周血细胞中的猫冠状病毒mRNA呈阳性;健康猫中,5%检测呈阳性。在一组81只经尸检确诊为FIP的动物中,75只(93%)检测呈阳性,而17只患有不同病症(非FIP病例)的猫均检测为阴性。鉴于假阳性结果率较低(高特异性),mRNA RT-PCR可能是FIP诊断手段中的一项有价值的补充。