Abstract：
Feline calicivirus (FCV) has a single-stranded, positive-sense RNA genome, and it is responsible for many infectious respiratory diseases in cats. In addition, more worryingly, highly virulent strains of FCV can cause high mortality in felines. Therefore, a rapid and reliable diagnosis tool plays an important role in controlling the outbreak of FCV. In this study, enzymatic recombinase amplification (ERA) assay combined with lateral flow dipstick (LFD) was developed for the detection of FCV, targeting a relatively conversed position of FCV-ORF1. The results showed that the optimal reaction condition was at 40 °C for 30 min. ERA-LFD method was highly sensitive with the detection limit as low as 3.2 TCID50 of FCV RNA per reaction. The specificity analysis demonstrated no cross-reactivity with feline parvovirus (FPV), feline herpesvirus (FHV) and feline infectious peritonitis virus (FIPV). ERA-LFD was highly repeatable and reproducible, with the intra-assay and inter-assay coefficients of variation for this method both less than 7%. The general test showed that all the recombinant plasmids with known mutant sites and FCV strains with different mutant sites stored in our laboratory were all detected by this method. Of the 23 samples, 14 samples were tested positive for FCV by ERA-LFD and RT-qPCR, respectively. In summary, ERA-LFD assay was a fast, accurate and convenient diagnosis tool for the detection of FCV. KEY POINTS: • The detection principle of ERA-LFD was introduced. • Almost all the currently known FCV strains can be detected. • ERA-LFD is easy to operate and can be used for field detection.
摘要：
猫杯状病毒（FCV）具有单链，正义RNA基因组，它是猫的许多传染性呼吸道疾病的原因。此外,更令人担忧的是,FCV的高毒力菌株可导致猫科动物的高死亡率。因此,快速可靠的诊断工具在控制FCV爆发中起着重要作用.在这项研究中,酶促重组酶扩增(ERA)结合侧流试纸(LFD)检测FCV,靶向FCV-ORF1的相对转换位置。结果表明，最佳反应条件为40℃反应30min。ERA-LFD方法灵敏度高，每个反应的FCVRNA的检出限低至3.2TCID50。特异性分析表明与猫细小病毒（FPV）没有交叉反应，猫疱疹病毒（FHV）和猫传染性腹膜炎病毒（FIPV）。ERA-LFD具有高度可重复性和可重复性，该方法的测定内和测定间变异系数均小于7%。一般测试表明，本实验室保存的所有具有已知突变位点的重组质粒和具有不同突变位点的FCV菌株均通过该方法检测到。在23个样本中，14份样本通过ERA-LFD和RT-qPCR检测为FCV阳性，分别。总之,ERA-LFD检测是一种快速的，准确，方便的诊断工具，用于FCV的检测。关键点:•介绍了ERA-LFD的检测原理。•几乎所有目前已知的FCV菌株都可以被检测到。•ERA-LFD易于操作，可用于现场检测。
