Development and evaluation of a point‐of‐care test with a combination of EZ‐Fast DNA extraction and real‐time PCR and LAMP detection: evaluation using blood samples containing the bovine leukaemia DNA
Development and evaluation of a point‐of‐care test with a combination of EZ‐Fast DNA extraction and real‐time PCR and LAMP detection: evaluation using blood samples containing the bovine leukaemia DNA
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DOI：
10.1111/lam.13376
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发表时间：
2020-08
影响因子：
2.4
通讯作者：
Y. Yamazaki;Uraiwan Thongchankaew-Seo;Konomu Nagao;H. Mekata;W. Yamazaki
中科院分区：
文献类型：
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作者：
Y. Yamazaki;Uraiwan Thongchankaew-Seo;Konomu Nagao;H. Mekata;W. Yamazaki
沿着社会全球化的进步，传染病的传播在世界范围内加速。部署高度敏感的基因检测对于早期诊断和早期遏制潜在的爆发和流行病以及常规监测至关重要，尽管繁琐和昂贵的核酸提取步骤是一个主要缺点。我们开发了一种简单、快速的DNA提取方法，命名为EZ‐Fast试剂盒，适用于现场环境。该试剂盒不需要先进的实验室设备或昂贵的DNA提取试剂盒，并且以极低的成本在10分钟内实现粗DNA提取，并且可以在现场设置中容易地进行。当结合真实的时间PCR和LAMP分析时，使用183份牛血液样本的POCT的性能与现有DNA提取方法的性能相似：92.5%（135/146）（真实的时间PCR）和93.7%（133/142）（LAMP）诊断灵敏度和100%诊断特异性。开发的POCT提供了一个强大的工具，以促进现场诊断在现场设置。
Along with progress in globalization of society, the spread of infectious diseases has accelerated worldwide. The deployment of highly sensitive genetic tests is essential for early diagnosis and early containment of potential outbreaks and epidemics, as well as routine surveillance, although tedious and expensive nucleic acid extraction steps represent a major drawback. Here we developed a simple and rapid DNA extraction method, named as an EZ‐Fast kit, applicable to the field setting. The kit does not require advanced laboratory equipment or expensive DNA extraction kits and achieves crude DNA extraction within 10 min at extremely low cost and can easily be performed in field settings. When combined with real‐time PCR and LAMP analyses, the performance of the POCT, using 183 bovine blood samples, was similar to that of the existing DNA extraction method: 92·5% (135/146) (real‐time PCR) and 93·7% (133/142) (LAMP) diagnostic sensitivities, and 100% diagnostic specificities. The developed POCT provides a powerful tool to facilitate on‐site diagnosis in a field setting.