Since both feline parvovirus (FPV) and feline bocavirus (FBoV) can cause diarrhea in cats, it is difficult to distinguish them clinically. This study aimed to develop a SYBR Green I-based duplex real-time polymerase chain reaction (PCR) assay for distinguishing FPV and FBoV-1 on the basis of the melting temperature of the PCR product. A total of 132 fecal samples from different domestic and feral cats were collected, and the results of SYBR Green I-based duplex real-time PCR assay were compared with those of the traditional PCR assay for a comprehensive evaluation. The melting temperatures were found to be 86 °C and 77.5 °C for FBoV-1 and FPV, respectively, and no specific melting peaks for other non-targeted feline viruses were observed. The data obtained from this assay had a good linear relationship; the detection limits of FPV and FBoV-1 were 2.907 × 10¹ copies/μL and 3.836 × 10¹ copies/μL, respectively. In addition, the experiment exhibited high reproducibility. The positive detection rates of the SYBR Green I-based duplex real-time PCR assay for FPV and FBoV-1 were 16.67% (22/132) and 6.82% (9/132), respectively, and the positive detection rate for co-infection with FPV and FBoV-1 was 3.03% (4/132). This result was much more sensitive than that of the traditional PCR method. Thus, the developed SYBR Green I-based assay is a sensitive, rapid, specific, and reliable method for the clinical diagnosis of FPV and FBoV-1 and can provide technical support for the simultaneous detection of co-infection with these viruses in the future.

由于猫细小病毒 (FPV) 和猫博卡病毒 (FBoV) 均可引起猫腹泻，因此临床上难以区分。本研究旨在开发一种基于 SYBR Green I 的双链实时聚合酶链反应 (PCR) 检测方法，用于根据 PCR 产物的解链温度区分 FPV 和 FBoV-1。共收集了来自不同家猫和野猫的 132 份粪便样本，将基于 SYBR Green I 的双工实时 PCR 检测结果与传统 PCR 检测结果进行比较，以进行综合评价。FBoV-1 和 FPV 的解链温度分别为 86°C 和 77.5°C，并且没有观察到其他非靶向猫科病毒的特定解链峰。本试验所得数据具有良好的线性关系；^分别为1拷贝/μL 和 3.836 × 10 ¹拷贝/μL。此外，该实验表现出很高的重现性。基于 SYBR Green I 的双工实时 PCR 检测 FPV 和 FBoV-1 的阳性检出率分别为 16.67% (22/132) 和 6.82% (9/132)，co- FPV 和 FBoV-1 感染率为 3.03% (4/132)。该结果比传统的 PCR 方法灵敏得多。因此，所开发的基于SYBR Green I的检测方法是临床诊断FPV和FBoV-1的一种灵敏、快速、特异性和可靠的方法，可为未来同时检测这些病毒的共感染提供技术支持。 .