Feline coronavirus (FCoV) contains two serotypes, feline enteric coronavirus (FECV) and Feline infectious peritonitis virus (FIPV). FECV and feline parvovirus (FPV) can cause similar clinical symptoms in cats, such as diarrhea. The objective of this study was to establish a duplex SYBR Green I-based quantitative polymerase chain reaction (qPCR) assay for rapid and simultaneous detection of FPV and FCoV. Two pairs of specific PCR primers were designed to target fragments of the VP2 gene of FPV and of the 5′ UTR gene of FCoV, respectively. The assay distinguished between the two viruses based on the melting curves (melting temperatures 77.0 ± 0.5 °C [FPV] and 80.5 ± 0.5 °C [FCoV]). The minimum limits of FPV and FCoV detection were 4.74 × 10¹ copies/μL and 7.77 × 10¹ copies/μL, respectively. The assay showed excellent reproducibility and reliability, based on the mean coefficient of variation. In conclusion, this novel duplex SYBR Green I-based qPCR assay is sensitive and can specifically, reliably, and rapidly detect FPV and FCoV (co-)infections.

猫冠状病毒 (FCoV) 包含两种血清型，猫肠道冠状病毒 (FECV) 和猫传染性腹膜炎病毒 (FIPV)。FECV 和猫细小病毒 (FPV) 可在猫中引起类似的临床症状，例如腹泻。本研究的目的是建立一种基于双链 SYBR Green I 的定量聚合酶链反应 (qPCR) 检测方法，用于快速和同时检测 FPV 和 FCoV。设计了两对特异性 PCR 引物，分别靶向 FPV 的 VP2 基因和 FCoV 的 5' UTR 基因的片段。该测定根据熔解曲线（熔解温度 77.0 ± 0.5 °C [FPV] 和 80.5 ± 0.5 °C [FCoV]）区分了两种病毒。FPV 和 FCoV 检测的最低限度为 4.74 × 10 ¹拷贝/μL 和 7.77 × 10 ¹拷贝/μL，分别。基于平均变异系数，该测定显示出极好的重现性和可靠性。总之，这种基于 SYBR Green I 的新型双链 qPCR 检测灵敏，可以特异性、可靠且快速地检测 FPV 和 FCoV（共）感染。