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 leukemia DNA,Letters in Applied Microbiology

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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 leukemia DNA
Letters in Applied Microbiology ( IF 2.0 ) Pub Date : 2020-09-23 , DOI: 10.1111/lam.13376
Y Yamazaki ₁ , U Thongchankaew-Seo ₁ , K Nagao ₂ , H Mekata ₃ , W Yamazaki _{1,

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Affiliation

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.

中文翻译：

EZ-Fast DNA 提取与实时 PCR 和 LAMP 检测相结合的床旁检测的开发和评估：使用含有牛白血病 DNA 的血液样本进行评估

随着社会全球化的发展，传染病在世界范围内的传播速度加快。尽管繁琐且昂贵的核酸提取步骤是一个主要缺点，但部署高度敏感的基因检测对于潜在爆发和流行病的早期诊断和早期遏制以及常规监测至关重要。在这里，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，适用于现场设置。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，可在 10 分钟内以极低的成本实现粗 DNA 提取，并且可以轻松地在现场环境中进行。当结合实时 PCR 和 LAMP 分析时，POCT 的性能，使用 183 个牛血样本，与现有 DNA 提取方法相似：92·5% (135/146) (实时 PCR) 和 93·7% (133/142) (LAMP) 诊断灵敏度，以及 100% 诊断特异性。开发的 POCT 提供了一个强大的工具来促进现场环境中的现场诊断。

更新日期：2020-09-23

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