College of Animal Science and Technology, Shandong Agricultural University, Tai'an, Shandong Province 271018, China; Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Tai'an, Shandong 271018, China; Shandong Provincial Engineering Technology Research Center of Animal Disease Control and Prevention, Tai'an, Shandong 271018, China.
College of Life Science, Qufu Normal University, Qufu, Shandong Province, China.
Poult Sci. 2020 Dec;99(12):6586-6592. doi: 10.1016/j.psj.2020.09.077. Epub 2020 Oct 8.
In November 2017, a severe infectious disease that devastated the major goose-producing regions in China was found to be caused by a novel goose astrovirus (N-AstV). The objective of this study was to develop a quantitative loop-mediated isothermal amplification (qLAMP) assay for the rapid diagnosis of N-AstV characterized with gout, hemorrhage, and swellings of the kidneys. A set of 4 specific primers, 2 inner and 2 outer primers, targeting the ORF1a gene of N-AstV were designed for the assay which could be completed within 60 min at 65°C in a water bath or on a real-time PCR instrument for quantitative analysis. The qLAMP assay showed a high sensitivity with a detection limit of 1 × 10 copies of the target DNA/μL. There were no cross-reactions with other viruses, and the reproducibility of the assay was confirmed in intrasensitivity and intersensitivity assay tests with variability ranging from 0.61 to 2.21%. The results indicated that the qLAMP assay for N-AstV was a simple, accurate, rapid, sensitive, and specific, especially useful for field detection.
2017 年 11 月，在中国主要鹅产区发现了一种严重的传染病，这种传染病是由一种新型鹅星状病毒（N-AstV）引起的。本研究旨在开发一种用于快速诊断具有痛风、出血和肾脏肿胀特征的 N-AstV 的定量环介导等温扩增（qLAMP）检测方法。该检测方法针对 N-AstV 的 ORF1a 基因设计了 4 对特异性引物，包括 2 对内引物和 2 对外引物，可在 65°C 的水浴或实时 PCR 仪器中在 60 分钟内完成，用于定量分析。qLAMP 检测方法具有很高的灵敏度，检测限为 1×10 拷贝目标 DNA/μL。与其他病毒无交叉反应，在灵敏度内和灵敏度间的重复性试验中得到了验证，变异系数范围为 0.61 至 2.21%。结果表明，N-AstV 的 qLAMP 检测方法简单、准确、快速、灵敏、特异性强，特别适用于现场检测。
