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 103 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.

中文翻译：

---

用于同时检测和区分番鸭细小病毒和鹅细小病毒的双重PCR检测方法。

番鸭细小病毒（MDPV）和鹅细小病毒（GPV）均可引起番鸭的高死亡率和发病率。MDPV和GPV具有较高的核苷酸同一性，这可能会导致在鉴别诊断时出错。在这项研究中，经过遗传比较后，选择了MDPVs和GPVs的NS基因来设计特异性引物。仅设计了三种引物（GF1，MF1和MGR1）用于双链PCR分析：GF1仅对GPV特异；MF1仅适用于MDPV；MGR1对于MDPV和GPV都高度保守。经过一系列优化实验后，双链PCR分析扩增了一个专门针对GPV的161 bp片段，一个专门针对MDPV的1197 bp片段，以及针对GPV和MDPV的两个片段（161 bp和1197 bp）。最低检测限为103拷贝/μl。使用来自番鸭中的其他病原体（包括DAdV-A，DuCV，DEV，GHPV，RA，大肠杆菌，PM和SS）的核酸无法获得扩增。在田间样品中应用双链PCR分析表明，即使是一天大的番鸭，MDPV阳性和GPV阳性也是如此。总之，仅使用三种高度特异性的引物就建立了同时检测和区分MDPV和GPV的双重PCR检测方法。我们的发现表明，在全国范围内对水禽接种MDPV和GPV疫苗是必要的。仅使用三种高度特异性的引物建立了用于同时检测和区分MDPV和GPV的双链PCR检测方法。我们的发现表明，在全国范围内对水禽接种MDPV和GPV疫苗是必要的。仅使用三种高度特异性的引物建立了用于同时检测和区分MDPV和GPV的双链PCR检测方法。我们的发现表明，在全国范围内对水禽接种MDPV和GPV疫苗是必要的。