Doki Tomoyoshi, Ohno Misaki, Kaku Mihoko, Odani Saori, To Kaito, Takano Tomomi
Laboratory of Veterinary Infectious Disease, School of Veterinary Medicine, Kitasato University, Towada, Aomori 034-8628, Japan.
Laboratory of Veterinary Infectious Disease, School of Veterinary Medicine, Kitasato University, Towada, Aomori 034-8628, Japan.
J Virol Methods. 2025 Jun 28;338:115214. doi: 10.1016/j.jviromet.2025.115214.
Feline infectious peritonitis (FIP) is a fatal disease in cats that is caused by feline coronavirus (FCoV). FCoV RT-qPCR is widely used to diagnose FIP due to its high sensitivity and ability to quantify FCoV RNA. However, its convenience is limited by the need for expensive equipment and/or processing at external laboratories. We herein developed two rapid and simple FCoV RNA detection systems: one combining conventional RT-PCR with the Single Tag Hybridization-Printed Array Strip (STH-PAS) method (the RT-PCR and STH-PAS system) and the other combining RT-RPA, an isothermal nucleic acid amplification method, with STH-PAS (the RT-RPA and STH-PAS system). Evaluations using FCoV RNA standards showed that the limit of detection for the RT-PCR and STH-PAS system was 10 copies/reaction, while that for the RT-RPA and STH-PAS system was 10 copies/reaction. The clinical performance of these systems was also examined using clinical samples from cats suspected of having FIP and compared to the conventional FCoV RT-qPCR genetic test. The results obtained showed a sensitivity of 66.7 % (95 % CI: 41.0-86.7) and specificity of 100 % (95 %CI: 9.4-100). These systems are a faster and simpler alternative to conventional methods, suggesting their potential in point-of-care testing in veterinary clinics.
猫传染性腹膜炎（FIP）是猫科动物中的一种致命疾病，由猫冠状病毒（FCoV）引起。FCoV RT-qPCR因其高灵敏度和定量FCoV RNA的能力而被广泛用于诊断FIP。然而，其便利性受到需要昂贵设备和/或在外部实验室进行处理的限制。我们在此开发了两种快速简单的FCoV RNA检测系统：一种将传统RT-PCR与单标签杂交印刷阵列条（STH-PAS）方法相结合（RT-PCR和STH-PAS系统），另一种将等温核酸扩增方法RT-RPA与STH-PAS相结合（RT-RPA和STH-PAS系统）。使用FCoV RNA标准品进行的评估表明，RT-PCR和STH-PAS系统的检测限为10拷贝/反应，而RT-RPA和STH-PAS系统的检测限为10拷贝/反应。还使用疑似患有FIP的猫的临床样本检查了这些系统的临床性能，并与传统的FCoV RT-qPCR基因检测进行了比较。获得的结果显示灵敏度为66.7%（95%CI：41.0-86.7），特异性为100%（95%CI：9.4-100）。这些系统是传统方法更快、更简单的替代方法，表明它们在兽医诊所即时检测中的潜力。
