Wang Chenchen, Liu Huixin, Cheng Junze, Pan Sijia, Yang Wenwen, Wei Xiaofang, Cheng Yue, Xu Ting, Si Hongbin
Guangxi Key Laboratory of Animal Breeding, Disease Control and Prevention, College of Animal Science and Technology, Guangxi Grass Station, Guangxi University, Nanning 530004, China.
Microorganisms. 2024 Nov 25;12(12):2423. doi: 10.3390/microorganisms12122423.
Duck Tembusu virus (DTMUV), duck hepatitis virus (DHV), Muscovy duck reovirus (MDRV), and Muscovy duck parvovirus (MDPV) represent four emergent infectious diseases impacting waterfowl, which can be challenging to differentiate due to overlapping clinical signs. In response to this, we have developed a one-step multiplex real-time fluorescence quantitative reverse transcription PCR (qRT-PCR) assay, capable of simultaneously detecting DTMUV, DHV, MDRV, and MDPV. This method exhibits high specificity, avoiding cross-reactivity with other viruses such as Fowl adenoviruses (FADV), infectious bursal disease virus (IBDV), infectious bronchitis virus (IBV), infectious laryngotracheitis virus (ILTV), Haemophilus paragallinarum (Hpg), duck circovirus (DUCV), goose astrovirus (GoAstV), and mycoplasma gallisepticum (MG). The limit of detection (LOD) established for DTMUV, DHV, MDRV, and MDPV was determined to be 27 copies/μL. In the repeatability test, the intra-assay and inter-assay coefficients of variation (CVs) of the recombinant plasmid standard were less than 2%. Utilizing this method, we analyzed 326 clinical specimens sourced from Guangxi over the period spanning October 2021 through December 2023, yielding promising and precise outcomes. The qRT-PCR method established herein exhibits commendable specificity, sensitivity, and repeatability. Furthermore, it boasts a high clinical detection rate, making it a highly effective tool for diagnosing these pathogenic agents in waterfowl.
鸭坦布苏病毒（DTMUV）、鸭肝炎病毒（DHV）、番鸭呼肠孤病毒（MDRV）和番鸭细小病毒（MDPV）是影响水禽的四种新发传染病，由于临床症状重叠，可能难以区分。针对这一情况，我们开发了一种一步多重实时荧光定量逆转录PCR（qRT-PCR）检测方法，能够同时检测DTMUV、DHV、MDRV和MDPV。该方法具有高特异性，避免了与其他病毒如禽腺病毒（FADV）、传染性法氏囊病病毒（IBDV）、传染性支气管炎病毒（IBV）、传染性喉气管炎病毒（ILTV）、副鸡嗜血杆菌（Hpg）、鸭圆环病毒（DUCV）、鹅星状病毒（GoAstV）和鸡毒支原体（MG）的交叉反应。确定DTMUV、DHV、MDRV和MDPV的检测限（LOD）为27拷贝/μL。在重复性试验中，重组质粒标准品的批内和批间变异系数（CVs）均小于2%。利用该方法，我们分析了2021年10月至2023年12月期间来自广西的326份临床样本，取得了良好而精确的结果。本文建立的qRT-PCR方法具有良好的特异性、敏感性和重复性。此外，它具有较高的临床检出率，是诊断水禽中这些病原体的高效工具。
