Anhui Province Key Laboratory of Veterinary Pathobiology and Disease Control, College of Animal Science and Technology, Anhui Agricultural University, Hefei, 230036, PR China.
Anhui Provincial Center for Animal Disease Control and Prevention, Hefei, 230000, PR China.
Mol Cell Probes. 2020 Oct;53:101648. doi: 10.1016/j.mcp.2020.101648. Epub 2020 Aug 14.
Beak atrophy and dwarfism syndrome (BADS) is commonly caused by co-infection with duck circovirus (DuCV) and novel goose parvovirus (NGPV). Therefore, concurrent detection of both viruses is important for monitoring and limiting BADS, although such a diagnostic test has not been reported. In this study, we developed a duplex, SYBR Green I-based real-time polymerase chain reaction (PCR) assay to enable the simultaneous detection of DuCV and NGPV. The assay readily distinguished between the two viruses, based on their different melting temperatures (Tm), where the Tm for DuCV was 80 °C and that for NGPV was 84.5 °C. Other non-target duck viruses that were tested did not show melting peaks. The detection limit of the duplex assay was 10 copies/μL for both viruses. This method exhibited high repeatability and reproducibility, and both the inter-assay and intra-assay variation coefficients were <1.6%. Thirty-one fecal samples were collected for clinical testing using real-time PCR analysis, and the results were confirmed using sequencing. The rate of co-infection was 6.5%, which was consistent with the sequencing results. This duplex real-time PCR assay offers advantages over other tests, such as rapid, sensitive, specific, and reliable detection of both viruses in a single sample, which enables the quantitative detection of DuCV and NGPV in clinical samples. Using this test may be instrumental in reducing the incidence of BADS and the associated economic losses in the duck and goose industries.
喙萎缩和矮小综合征（BADS）通常由鸭圆环病毒（DuCV）和新型鹅细小病毒（NGPV）的合并感染引起。因此，同时检测这两种病毒对于监测和限制 BADS 非常重要，尽管尚未报道这种诊断测试。在这项研究中，我们开发了一种基于双重 SYBR Green I 的实时聚合酶链反应（PCR）检测方法，能够同时检测 DuCV 和 NGPV。该检测方法基于两种病毒不同的熔点（Tm）来区分它们，DuCV 的 Tm 为 80°C，NGPV 的 Tm 为 84.5°C。其他测试的非目标鸭病毒没有出现熔解峰。该双重检测方法的检测限对两种病毒均为 10 拷贝/μL。该方法表现出高度的重复性和再现性，并且组间和组内变异系数均<1.6%。使用实时 PCR 分析对 31 份粪便样本进行了临床检测，并用测序进行了验证。合并感染率为 6.5%，与测序结果一致。与其他测试相比，该双重实时 PCR 检测方法具有优势，例如在单个样品中快速、敏感、特异性和可靠地同时检测两种病毒，并能够对临床样本中的 DuCV 和 NGPV 进行定量检测。使用该测试可能有助于降低鸭和鹅产业中 BADS 的发病率和相关经济损失。
