Institute of Animal Husbandry and Veterinary Medicine of Fujian Academy of Agricultural Sciences, Fuzhou, 350013, China.
Fujian Provincial Key Laboratory for Avian Diseases Control and Prevention & Fujian Animal Diseases Control Technology Development Center, Fuzhou, 350013, China.
BMC Vet Res. 2019 Nov 1;15(1):389. doi: 10.1186/s12917-019-2090-7.
Classic goose parvovirus (cGPV) causes high mortality and morbidity in goslings and Muscovy ducklings. Novel GPV (N-GPV) causes short beak and dwarfism syndrome (SBDS) in Cherry Valley ducks, Pekin ducks and Mule ducks. Both cGPV and N-GPV have relatively strict host specificity, with obvious differences in pathogenicity. Specific detection of cGPV and N-GPV may result in false positives due to high nucleotide similarity with Muscovy duck parvovirus (MDPV). The aim of this study was to develop a highly specific, sensitive, and reliable TaqMan real-time PCR (TaqMan qPCR) assay for facilitating the molecular detection of cGPV and N-GPV.
After genetic comparison, the specific conserved region (located on the NS gene) of cGPV and N-GPV was selected for primer and probe design. The selected regions were significantly different from MDPV. Through a series of optimization experiments, the limit of detection was 50.2 copies/μl. The assay was highly specific for the detection of cGPV and N-GPV and no cross-reactivity was observed with E. coli., P.M., R.A., S.S., MDPV, N-MDPV, DAdV-A, DEV, GHPV, DHAV-1, DHAV-3, ATmV, AIV, MDRV and N-DRV. The assay was reproducible with an intra-assay and inter-assay variability of less than 2.37%. Combined with host specificity, the developed TaqMan qPCR can be used for cGPV and N-GPV in differential diagnoses. The frequency of cGPV in Muscovy duckling and goslings was determined to be 12 to 44%, while N-GPV frequency in Mule ducks and Cherry Valley ducks was 36 to 56%. Additionally, fluorescence-positive signals can be found in Mule duck embryos and newly hatched Mule ducklings. These findings provide evidence of possible vertical transmission of N-GPV from breeding Mule ducks to ducklings.
We established a quantitative platform for epidemiological investigations and pathogenesis studies of cGPV and N-GPV DNA that was highly sensitive, specific, and reproducible. N-GPV and cGPV infections can be distinguished based on host specificity.
经典鹅细小病毒（cGPV）可导致鹅苗和麝香鸭苗高死亡率和高发病率。新型鹅细小病毒（N-GPV）可导致樱桃谷鸭、北京鸭和骡鸭发生短喙矮小综合征（SBDS）。cGPV 和 N-GPV 均具有相对严格的宿主特异性，其致病性存在明显差异。由于与麝香鸭细小病毒（MDPV）的核苷酸相似度较高，因此对 cGPV 和 N-GPV 的特异性检测可能会导致假阳性。本研究旨在开发一种高度特异、敏感、可靠的 TaqMan 实时 PCR（TaqMan qPCR）检测方法，以促进 cGPV 和 N-GPV 的分子检测。
通过遗传比较，选择 cGPV 和 N-GPV 的特定保守区域（位于 NS 基因上）进行引物和探针设计。所选区域与 MDPV 有明显差异。通过一系列优化实验，检测限为 50.2 拷贝/μl。该检测方法对 cGPV 和 N-GPV 的检测具有高度特异性，与大肠杆菌、P.M.、R.A.、S.S.、MDPV、N-MDPV、DAdV-A、DEV、GHPV、DHAV-1、DHAV-3、ATmV、AIV、MDRV 和 N-DRV 无交叉反应。该检测方法具有良好的重复性，内、间试验变异系数均小于 2.37%。结合宿主特异性，建立的 TaqMan qPCR 可用于 cGPV 和 N-GPV 的鉴别诊断。检测到雏鹅和雏麝香鸭中 cGPV 的频率为 12%至 44%，而骡鸭和樱桃谷鸭中 N-GPV 的频率为 36%至 56%。此外，在骡鸭胚胎和刚孵化的骡鸭中可检测到荧光阳性信号。这些发现为 N-GPV 从种骡鸭垂直传播到雏鸭提供了证据。
我们建立了一种用于 cGPV 和 N-GPV DNA 流行病学调查和发病机制研究的定量平台，该平台具有高度的敏感性、特异性和重复性。根据宿主特异性可区分 N-GPV 和 cGPV 感染。
