State Key Laboratory of Veterinary Etiological Biology and National Foot-and-Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou, Gansu, 730046, PR China.
State Key Laboratory of Veterinary Etiological Biology and National Foot-and-Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou, Gansu, 730046, PR China; College of Animal Science, Yangtze University, Jingmi Street, Jingzhou District, Jingzhou, 434025, PR China.
Mol Cell Probes. 2020 Oct;53:101643. doi: 10.1016/j.mcp.2020.101643. Epub 2020 Aug 5.
Porcine vesicular disease caused by Senecavirus A (SVA) is a newly emerging disease in many countries. Based on clinical signs only, it is very challenging to distinguish SVA infection from other similar diseases, such as foot and mouth disease, swine vesicular disease, and vesicular stomatitis. Therefore, it is crucial to establish a detection assay for the clinical diagnosis of SVA infection. In this study, a pair of specific primers were designed based on the highly conserved L/VP4 gene sequence of SVA. The established SYBR green I-based quantitative reverse transcription polymerase chain reaction (qRT-PCR) method was used to detect SVA nucleic acids in clinical samples. The limit of detection SVA nucleic acids by qRT-PCR was 6.4 × 10 copies/μL, which was significantly more sensitive than that by gel electrophoresis of 6.4 × 10 copes/μL. This assay was specific and had no cross-reaction with other seven swine viruses. Using SYBR green I-based qRT-PCR, the SVA positive rates in experimental animal samples and field samples were 67.60% (96/142) and 80% (24/30) respectively. The results demonstrate that SYBR green I-based qRT-PCR is a rapid and specific method for the clinical diagnosis and epidemiological investigation of related vesicular diseases caused by SVA.
猪水泡病由 Senecavirus A（SVA）引起，是许多国家新出现的疾病。仅根据临床症状，很难将 SVA 感染与其他类似疾病（如口蹄疫、猪水泡病和水疱性口炎）区分开来。因此，建立一种用于 SVA 感染临床诊断的检测方法至关重要。本研究基于 SVA 的高度保守 L/VP4 基因序列设计了一对特异性引物。建立的 SYBR green I 实时荧光定量 RT-PCR（qRT-PCR）方法用于检测临床样本中的 SVA 核酸。qRT-PCR 检测 SVA 核酸的灵敏度为 6.4×10 拷贝/μL，明显高于凝胶电泳的 6.4×10 拷贝/μL。该检测方法具有特异性，与其他七种猪病毒无交叉反应。使用 SYBR green I 实时荧光定量 RT-PCR，实验动物样本和现场样本中的 SVA 阳性率分别为 67.60%（96/142）和 80%（24/30）。结果表明，SYBR green I 实时荧光定量 RT-PCR 是一种快速、特异的用于 SVA 引起的相关水泡病的临床诊断和流行病学调查的方法。
