Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agriculture Sciences, Fuzhou, China; Fujian Animal Diseases Control Technology Development Center, Fujian Academy of Agriculture Sciences, Fuzhou, China.
Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agriculture Sciences, Fuzhou, China.
Poult Sci. 2021 Feb;100(2):482-487. doi: 10.1016/j.psj.2020.11.016. Epub 2020 Nov 19.
H146-like goose-origin calicivirus (H146-like GCV) is a novel Caliciviridae family member in the Sanovirus genus that was recently discovered and proposed to cause runting-stunting syndrome and urate deposition in geese. At present, however, there is a lack of epidemiological information pertaining to the dynamics and distribution of H146-like GCV. The development of novel molecular diagnostic approaches capable of rapidly and accurately detecting this virus would support the strengthening, the prevention, and control of H146-like GCV infection. In the present study, we therefore used a TaqMan probe and primers specific for the viral nonstructural (NS) gene to develop a highly sensitive and specific PCR assay capable of detecting this H146-like GCV. The assay reproducibly detected 5.07 × 10 copies of a recombinant DNA plasmid containing the NS gene, with a dynamic range of 8 orders of magnitude (10-10 copies). Importantly, no cross-reactivity was observed with common viruses that affected waterfowl, and when we used this assay to evaluate clinical samples, we found it to be more sensitive and faster than traditional PCR. In summary, herein, we developed a novel TaqMan-based real-time PCR approach that could reliably detect and diagnose H146-like GCV. This tool will allow for the real-time diagnosis of H146-like GCV infections, enabling researchers to better understand the epidemiology and clinical presentation of this disease.
类 H146 鹅源杯状病毒（H146-like GCV）是最近发现的新型杯状病毒科沙诺病毒属成员，被提议引起鹅的矮小综合征和尿酸沉积。然而，目前缺乏关于 H146-like GCV 的动态和分布的流行病学信息。开发能够快速准确检测这种病毒的新型分子诊断方法将支持加强、预防和控制 H146-like GCV 感染。在本研究中，我们使用针对病毒非结构（NS）基因的 TaqMan 探针和引物，开发了一种高度敏感和特异的 PCR 检测方法，能够检测这种 H146-like GCV。该检测方法可重复性地检测到含有 NS 基因的重组 DNA 质粒的 5.07×10 拷贝，动态范围为 8 个数量级（10-10 拷贝）。重要的是，与影响水禽的常见病毒没有交叉反应性，当我们使用该检测方法评估临床样本时，发现其比传统 PCR 更敏感和快速。总之，我们开发了一种新的基于 TaqMan 的实时 PCR 方法，能够可靠地检测和诊断 H146-like GCV。该工具将能够实时诊断 H146-like GCV 感染，使研究人员能够更好地了解这种疾病的流行病学和临床表现。
