环介导等温扩增检测技术用于快速可视化检测新型猪圆环病毒 3。
Loop-mediated isothermal amplification assay for the rapid and visual detection of novel porcine circovirus 3.
机构信息
College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University, Daegu 41566, Republic of Korea.
Animal Disease Diagnostic Division, Animal and Plant Quarantine Agency, Gimcheon 39660, Republic of Korea.
出版信息
J Virol Methods. 2018 Mar;253:26-30. doi: 10.1016/j.jviromet.2017.12.006. Epub 2017 Dec 24.
A loop-mediated isothermal amplification (LAMP) assay using hydroxynaphthol blue was developed for the rapid and visual detection of the capsid gene of porcine circovirus 3 (PCV3). The amplification could be completed in 40 min at 62°C, and the results could be visually detected by the naked eye. The assay specifically amplified PCV3 DNA and not other porcine viral nucleic acids. The limit of detection of the assay was 50 PCV3 DNA copies, which was comparable to that of the real-time polymerase chain reaction (qPCR) and lower than that of conventional PCR. In the clinical evaluation, the PCV3 detection rate of the LAMP assay was higher than that of PCR and agreed 100% with that of qPCR. These results indicate that the LAMP assay will be a valuable tool for the rapid, sensitive, and specific detection of PCV3 in clinical samples.
建立了一种使用羟基萘酚蓝的环介导等温扩增（LAMP）检测方法，用于快速和可视化检测猪圆环病毒 3（PCV3）的衣壳基因。该扩增可在 62°C 下 40 分钟内完成，结果可通过肉眼直接观察。该检测方法特异性扩增 PCV3 DNA，而不扩增其他猪病毒核酸。该检测方法的检测限为 50 个 PCV3 DNA 拷贝，与实时聚合酶链反应（qPCR）相当，低于常规 PCR。在临床评估中，LAMP 检测方法的 PCV3 检测率高于 PCR，与 qPCR 完全一致，达到 100%。这些结果表明，LAMP 检测方法将成为快速、敏感、特异性检测临床样本中 PCV3 的有价值的工具。
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