Zhongkai University of Agriculture and Engineering, Guangdong, Guangzhou, 510225, China; Guangdong Province Key Laboratory of Waterfowl Healthy Breeding, Guangdong, Guangzhou, 510225, China.
Foshan University, Guangdong, Foshan, 528000, China.
J Virol Methods. 2019 Apr;266:34-40. doi: 10.1016/j.jviromet.2019.01.010. Epub 2019 Jan 22.
Goose parvovirus (GPV) is one of the most serious viral pathogens in goslings. Recently, a new pathogen to the Chinese mainland-duck-origin novel goose parvovirus (N-GPV)-was found to be 90.8-94.6% identical to the nucleotide sequence of GPV, and typically causes growth disorders and high infection rates in meat ducks. The spread of both of these viruses hinders the healthy development of the waterfowl breeding industry. In this study, recombinase polymerase amplification (RPA) was combined with a vertical flow (VF) visualization strip to develop a universal assay for the rapid detection of GPV and N-GPV. A set of specific primers and probes were designed to target the VP3 gene. Detection was possible at a constant temperature of 37 °C within 5-10 min. The assay successfully detected GPV and N-GPV with high-specificity and did not exhibit cross-reactivity with other waterfowl viruses and bacteria. The analytical sensitivity of the GPV-RPA-VF assay was 2 × 10 copies of GPV plasmid. Validation of the GPV-RPA-VF assay-using 60 samples from the field--confirmed 100% similarity between the results of GPV-RPA-VF and conventional qPCR. The results indicate that the GPV-RPA-VF assay was accurate, sensitive, and specific. This assay can be performed with minimal equipment and training to rapidly detect GPV and N-GPV during the early phase of an outbreak, especially when timely veterinary diagnoses are needed in the field and in rural areas.
鹅细小病毒（GPV）是小鹅中最严重的病毒病原体之一。最近，一种新的病原体——鸭源新型鹅细小病毒（N-GPV）被发现与 GPV 的核苷酸序列相似度为 90.8-94.6%，通常会导致肉鸭生长障碍和高感染率。这两种病毒的传播阻碍了水禽养殖业的健康发展。在本研究中，重组酶聚合酶扩增（RPA）与垂直流（VF）可视化条结合，开发了一种用于快速检测 GPV 和 N-GPV 的通用检测方法。设计了一组针对 VP3 基因的特异性引物和探针。在 37°C 的恒温下，检测可以在 5-10 分钟内完成。该检测方法对 GPV 和 N-GPV 具有高度特异性，与其他水禽病毒和细菌无交叉反应。GPV-RPA-VF 检测方法的分析灵敏度为 2×10 拷贝的 GPV 质粒。使用来自现场的 60 个样本对 GPV-RPA-VF 检测方法进行验证，结果表明 GPV-RPA-VF 和常规 qPCR 的结果之间具有 100%的相似性。结果表明，GPV-RPA-VF 检测方法准确、敏感且具有特异性。该检测方法可在最低限度的设备和培训下进行，可在疫情早期快速检测 GPV 和 N-GPV，特别是在现场和农村地区需要及时兽医诊断时。
