Development of RT-LAMP and real-time RT-PCR assays for the rapid detection of the new duck Tembusu-like BYD virus
Development of RT-LAMP and real-time RT-PCR assays for the rapid detection of the new duck Tembusu-like BYD virus
复制标题
开发 RT-LAMP 和实时 RT-PCR 检测方法，用于快速检测新型鸭 Tembusu 样 BYD 病毒
DOI：
10.1007/s00705-012-1431-7
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发表时间：
2012-12-01
影响因子：
2.7
通讯作者：
Qin, Cheng-Feng
中科院分区：
文献类型：
--
作者：
Jiang, Tao;Liu, Juan;Qin, Cheng-Feng
一种新的鸭坦布苏病毒（TMUV），也被称为比亚迪病毒，已被确定为中国大陆新出现的鸭掉蛋综合征的病原体。新型TMUV在中国大陆的迅速传播和广泛分布，给家禽业造成了重大损失，并对公众健康构成了巨大威胁。快速和灵敏的检测方法对于预防和控制TMUV感染至关重要。在这项研究中，我们分别开发了一种针对鸭TMUV的逆转录环介导的等温扩增方法（RT-LAMP）和一种基于SYBR green - i的实时RT-PCR方法，并在实验室和现场样品中进行了验证。RT-LAMP法和实时RT-PCR法每个反应的检出限分别为1 × 10−4和1 × 10−3PFU。与黄病毒属其他相关成员进行特异性分析，未发现交叉反应。此外，这两种分析方法都用现场样品进行了评估，它们表现出高灵敏度和特异性。此外，实时RT-PCR检测在病毒载量分析中效果良好，这表明脾脏可能是鸭TMUV在鸭体内复制的主要靶点。TMUV特异性RT-LAMP和实时RT-PCR检测将为TMUV感染的诊断和流行病学监测提供有用的工具。
A new duck Tembusu virus (TMUV), also known as BYD virus, has been identified as the causative agent for the emerging duck egg-drop syndrome in mainland China. The rapid spread and wide distribution of the new TMUV in mainland China result in heavy loss to the poultry industry and pose great threats to public health. Rapid and sensitive detection methods are critical for prevention and control of TMUV infections. In this study, a reverse-transcription loop-mediated isothermal amplification assay (RT-LAMP) and an SYBR Green-I–based real-time RT-PCR assay specific for the duck TMUV were developed and validated with laboratory and field samples, respectively. The detection limits were 1 × 10−4and 1 × 10−3PFU per reaction for the RT-LAMP assay and real-time RT-PCR assay, respectively. The specificities were analyzed with other related members of the genusFlavivirus, and no cross-reaction was observed. Furthermore, both assays were evaluated with field samples, and they exhibited high sensitivity and specificity. In addition, the real-time RT-PCR assay worked well in viral load analysis, which revealed that the spleen may be the primary target for the replication of new duck TMUV in ducks. The TMUV-specific RT-LAMP and real-time RT-PCR assays will provide useful tools for the diagnosis and epidemiological surveillance of TMUV infection.