College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengdong New District Longzi Lake 15#, Zhengzhou, 450046, Henan Province, People's Republic of China.
College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengdong New District Longzi Lake 15#, Zhengzhou, 450046, Henan Province, People's Republic of China; Zhengzhou Major Pig Disease Prevention and Control Laboratory, Zhengzhou, 450046, Henan Province, People's Republic of China.
Mol Cell Probes. 2019 Apr;44:44-50. doi: 10.1016/j.mcp.2019.02.002. Epub 2019 Feb 5.
The development of a rapid, specific, and sensitive SYBR Green I-based duplex real-time quantitative PCR assay is described for the simultaneous detection of porcine epidemic diarrhea virus (PEDV) and porcine circovirus type 3 (PCV3). The assay specifically detected PEDV and PCV3, with no fluorescence detected for other non-targeted pig pathogens. The assay showed a good linear relationship, and the limits of detection for this assay were 34.6 copies/μL and 61.2 copies/μL for PEDV and PCV3, respectively. The assay exhibited high repeatability and reproducibility, with intra-assay and inter-assay variation coefficients less than 2.0%. A clinical evaluation using intestinal tissue and fecal samples from piglets suffering from diarrhea at different pig farms in China revealed that the singular infection rates of PEDV and PCV3 were 43.94% (29/66) and 16.67% (11/66), respectively, while the co-infection rate of PCV3 with PEDV was 27.27% (18/66). The results indicate this assay is a rapid and reliable diagnostic tool for PEDV and PCV3 monitoring and surveillance in the field, and provides technical support for the quantitative detection of clinical samples infected or co-infected with PEDV and PCV3.
本文描述了一种基于 SYBR Green I 的快速、特异、敏感的双重实时定量 PCR 检测方法，用于同时检测猪流行性腹泻病毒（PEDV）和猪圆环病毒 3 型（PCV3）。该方法特异性检测 PEDV 和 PCV3，对其他非目标猪病原体无荧光检测。该方法显示出良好的线性关系，PEDV 和 PCV3 的检测限分别为 34.6 拷贝/μL 和 61.2 拷贝/μL。该方法具有良好的重复性和再现性，内、间试验变异系数均小于 2.0%。对来自中国不同猪场腹泻仔猪的肠道组织和粪便样本进行临床评估表明，PEDV 和 PCV3 的单一感染率分别为 43.94%（29/66）和 16.67%（11/66），而 PCV3 与 PEDV 的共同感染率为 27.27%（18/66）。结果表明，该方法是一种快速可靠的 PEDV 和 PCV3 监测诊断工具，为 PEDV 和 PCV3 感染或共同感染的临床样本的定量检测提供了技术支持。
