Cao Nan, Tang Zhihui, Zhang Xiyu, Li Wanyan, Li Bingxin, Tian Yunbo, Xu Danning
Guangdong Province Key Laboratory of Waterfowl Healthy Breeding, College of Animal Science and Technology, Zhongkai University of Agriculture and Engineering, Guangzhou, China.
MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China.
Front Vet Sci. 2022 Feb 8;8:792322. doi: 10.3389/fvets.2021.792322. eCollection 2021.
As companion animals, felines play an important role in human's family life, and their healthcare has attracted great attention. Viruses such as feline calicivirus (FCV), feline herpesvirus 1 (FHV-1), and feline parvovirus virus (FPV) are the most common pathogens that cause severe infectious disease in baby cats. Thus, preclinical detection and intervention of these three viruses is an effective means to prevent diseases and minimize their danger condition. In this study, a triplex TaqMan quantitative real-time polymerase chain reaction (qRT-PCR) assay was developed to detect these three viruses simultaneously. The detection limit of FPV, FCV, and FHV-1 was 5 × 10 copies/assay, which exhibited higher sensitivity (about 10- to100-fold) than conventional PCR. The coefficients of variation (CVs) of the intra-assay variability were lower than 1.86%, and that of inter-assay variability were lower than 3.19%, indicating the excellent repeatability and reproducibility of the triplex assay. Additionally, the assay showed good specificity. Finally, samples from 48 cats were analyzed using the established assay and commercial kits. As a result, the total positive rates for these viruses were 70.83 or 62.5%, respectively, which demonstrated that the developed qRT-PCR assay was more accurate than the commercial kits and could be used in clinical diagnosis.
作为伴侣动物，猫科动物在人类家庭生活中扮演着重要角色，其医疗保健受到了极大关注。猫杯状病毒（FCV）、猫疱疹病毒1型（FHV-1）和猫细小病毒（FPV）等病毒是导致幼猫严重传染病的最常见病原体。因此，对这三种病毒进行临床前检测和干预是预防疾病并将其危险状况降至最低的有效手段。在本研究中，开发了一种三重TaqMan定量实时聚合酶链反应（qRT-PCR）检测方法，用于同时检测这三种病毒。FPV、FCV和FHV-1的检测限为5×10拷贝/检测，其灵敏度比传统PCR高（约10至100倍）。批内变异系数（CVs）低于1.86%，批间变异系数低于3.19%，表明该三重检测方法具有出色的重复性和再现性。此外，该检测方法具有良好的特异性。最后，使用建立的检测方法和商业试剂盒对48只猫的样本进行了分析。结果，这些病毒的总阳性率分别为70.83%或62.5%，这表明所开发的qRT-PCR检测方法比商业试剂盒更准确，可用于临床诊断。
