Fujian Provincial Key Laboratory for Avian Diseases Control and Prevention, Fujian Animal Diseases Control Technology Development Center, 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, Institute of Animal Husbandry and Veterinary Medicine of Fujian Academy of Agricultural Sciences, Fuzhou, 350013, China.
Mol Cell Probes. 2019 Oct;47:101439. doi: 10.1016/j.mcp.2019.101439. Epub 2019 Aug 21.
Both Muscovy duck parvovirus (MDPV) and goose parvovirus (GPV) can cause high mortality and morbidity in Muscovy ducklings. MDPVs and GPVs share high nucleotide identity, which can cause errors during differential diagnosis. In this study, the NS genes of both MDPVs and GPVs were chosen for the design of specific primers after genetic comparison. Only three primers (GF1, MF1 and MGR1) were designed for the duplex PCR assay: GF1 is specific for GPV only; MF1 is specific for MDPV only; and MGR1 is highly conserved for both MDPV and GPV. After a series of optimization experiments, the duplex PCR assay amplified a 161-bp fragment specifically for GPV, a 1197-bp fragment specifically for MDPV, and two fragments (161-bp and 1197-bp) for both GPV and MDPV. The lowest detection limit was 10 copies/μl. No amplification was obtained using nucleic acids from other pathogens (including DAdV-A, DuCV, DEV, GHPV, R.A., E. coli., P.M. and S.S.) occurring in Muscovy ducks. Application of the duplex PCR assay in field samples showed that even one-day-old Muscovy ducklings were both MDPV-positive and GPV-positive. In conclusion, a duplex PCR assay for the simultaneous detection and differentiation of MDPV and GPV was established using only three highly specific primers. Our finding suggested that country-wide vaccination with MDPV and GPV vaccines in waterfowls are necessary.
Muscovy duck parvovirus (MDPV) 和鹅细小病毒 (GPV) 均可导致雏番鸭发生高死亡率和高发病率。MDPV 和 GPV 具有高度的核苷酸同源性，这可能会导致在鉴别诊断过程中出现错误。在本研究中，通过遗传比较，选择 MDPV 和 GPV 的 NS 基因用于设计特异性引物。该双重 PCR 检测方法仅设计了 3 对引物（GF1、MF1 和 MGR1）：GF1 仅特异性针对 GPV；MF1 仅特异性针对 MDPV；而 MGR1 则对 MDPV 和 GPV 高度保守。经过一系列优化实验，该双重 PCR 检测方法特异性扩增出 161-bp 片段用于 GPV，1197-bp 片段用于 MDPV，以及同时用于 GPV 和 MDPV 的两个片段（161-bp 和 1197-bp）。最低检测限为 10 拷贝/μl。使用番鸭中其他病原体（包括 DAdV-A、DuCV、DEV、GHPV、R.A.、大肠杆菌、P.M. 和 S.S.）的核酸均未获得扩增。该双重 PCR 检测方法在田间样品中的应用表明，即使是 1 日龄的雏番鸭，也同时 MDPV 和 GPV 阳性。总之，建立了一种使用仅 3 对高度特异性引物同时检测和区分 MDPV 和 GPV 的双重 PCR 检测方法。我们的发现表明，在水禽中进行 MDPV 和 GPV 疫苗的全国性免疫接种是必要的。
