Longstaff Louise, Porter Emily, Crossley Victoria J, Hayhow Sophie E, Helps Christopher R, Tasker Séverine
1 The Feline Centre, Langford Veterinary Services and School of Veterinary Sciences, University of Bristol, Langford, Bristol, UK.
2 School of Veterinary Sciences, University of Bristol, Langford, Bristol, UK.
J Feline Med Surg. 2017 Feb;19(2):240-245. doi: 10.1177/1098612X15606957. Epub 2016 Jul 10.
Objectives The aim of the study was to determine whether feline coronavirus (FCoV) RNA in effusion samples can be used as a diagnostic marker of feline infectious peritonitis (FIP); and in FCoV RNA-positive samples to examine amino acid codons in the FCoV spike protein at positions 1058 and 1060 where leucine and alanine, respectively, have been associated with systemic or virulent (FIP) FCoV infection. Methods Total RNA was extracted from effusion samples from 20 cats with confirmed FIP and 23 cats with other diseases. Feline coronavirus RNA was detected using a reverse transcriptase quantitative polymerase chain reaction assay (qRT-PCR), and positive samples underwent pyrosequencing of position 1058 with or without Sanger sequencing of position 1060 in the FCoV spike protein. Results Seventeen (85%) of the effusion samples from 20 cats with FIP were positive for FCoV RNA, whereas none of the 23 cats with other diseases were positive. Pyrosequencing of the 17 FCoV-positive samples showed that 11 (65%) of the cats had leucine and two (12%) had methionine at position 1058. Of the latter two samples with methionine, one had alanine at position 1060. Conclusions and relevance A positive FCoV qRT-PCR result on effusions appears specific for FIP and may be a useful diagnostic marker for FIP in cats with effusions. The majority of FCoVs contained amino acid changes previously associated with systemic spread or virulence (FIP) of the virus.
目的 本研究旨在确定积液样本中的猫冠状病毒(FCoV)RNA是否可作为猫传染性腹膜炎(FIP)的诊断标志物;并对FCoV RNA阳性样本检测FCoV刺突蛋白中第1058和1060位的氨基酸密码子,其中亮氨酸和丙氨酸分别与全身性或强毒力(FIP)FCoV感染相关。方法 从20只确诊为FIP的猫和23只患有其他疾病的猫的积液样本中提取总RNA。使用逆转录定量聚合酶链反应检测法(qRT-PCR)检测猫冠状病毒RNA,对阳性样本进行FCoV刺突蛋白第1058位的焦磷酸测序,第1060位视情况进行桑格测序。结果 20只患有FIP的猫的积液样本中有17只(85%)FCoV RNA呈阳性,而23只患有其他疾病的猫均为阴性。对17份FCoV阳性样本进行焦磷酸测序显示,11只(65%)猫在第1058位为亮氨酸,2只(12%)为甲硫氨酸。在这2份含甲硫氨酸的样本中,有1份在第1060位为丙氨酸。结论及意义 积液样本FCoV qRT-PCR结果呈阳性似乎对FIP具有特异性,可能是积液性猫FIP的有用诊断标志物。大多数FCoV含有先前与病毒全身性传播或毒力(FIP)相关的氨基酸变化。