College of Animal Science, South China Agricultural University, Guangzhou 510642, People's Republic of China.
Poult Sci. 2010 Mar;89(3):477-83. doi: 10.3382/ps.2009-00527.
Muscovy duck parvovirus (MDPV) usually causes high morbidity and mortality in 1- to 3-wk-old Muscovy ducklings due to serious infections, which is an imminent threat to the commercial duck industry in China. The objectives of this study were to develop and evaluate a simple, rapid, and inexpensive loop-mediated isothermal amplification (LAMP) method for specific detection of MDPV and to compare it with the PCR method in rapidity, sensitivity, and accuracy. The novel LAMP assay used a set of 4 specific primers to recognize 6 distinct genomic sequences of capsid protein (VP3) from MDPV, which could be completed within 50 min at 63 degrees C in a simple water bath. The diagnostic results demonstrated that the LAMP assay detected all 7 preserved MDPV isolates, had no cross-reactivity with other duck pathogens (i.e., goose parvovirus, duck plague virus, H9N2 avian influenza virus, duck hepatitis type virus I, and Muscovy duck reovirus). The LAMP assay was at least 10-fold more sensitive than the routine PCR assay and obtained more sensitivity in 61 clinical samples. Therefore, the newly developed LAMP assay provides a specific and sensitive means for detecting MDPV and can be simply applied both in field conditions and in laboratory operations in a cost-effective manner with primary care facilities.
Muscovy 鸭细小病毒（Muscovy duck parvovirus，MDPV）通常会导致 1 至 3 周龄的麝香鸭雏鸭出现高发病率和死亡率，因为严重感染，这对中国商业鸭产业构成了迫在眉睫的威胁。本研究的目的是开发和评估一种简单、快速、廉价的环介导等温扩增（loop-mediated isothermal amplification，LAMP）方法，用于特异性检测 MDPV，并将其与 PCR 方法在快速性、敏感性和准确性方面进行比较。新的 LAMP 检测方法使用一组 4 个特异性引物来识别 MDPV 衣壳蛋白（VP3）的 6 个独特基因组序列，可在 63°C 的简单水浴中在 50 分钟内完成。诊断结果表明，LAMP 检测法可检测到所有 7 个保存的 MDPV 分离株，与其他鸭病原体（即鹅细小病毒、鸭瘟病毒、H9N2 禽流感病毒、鸭肝炎病毒 I 和麝香鸭呼肠孤病毒）无交叉反应。LAMP 检测法比常规 PCR 检测法至少灵敏 10 倍，在 61 份临床样本中获得了更高的灵敏度。因此，新开发的 LAMP 检测法为检测 MDPV 提供了一种特异性和灵敏性的方法，可以在现场条件下简单应用，并且在具有基本医疗设施的实验室操作中具有成本效益。
