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Biology
通过逆转录环介导的等温扩增检测 SARS-CoV-2 病毒
通过逆转录环介导的等温扩增检测 SARS-CoV-2 病毒
JoVE Journal
Biology
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JoVE Journal Biology
Detecting SARS-CoV-2 Virus by Reverse Transcription-Loop-Mediated Isothermal Amplification

通过逆转录环介导的等温扩增检测 SARS-CoV-2 病毒

Full Text
1,036 Views
05:34 min
September 8, 2023

DOI: 10.3791/65662-v

Steven A. David-Jimenez1, Paola A. Caicedo1, Maria F. Villegas-Torres1,2, Natalia Campillo-Pedroza1,3

1Facultad de Ingeniería, Diseño y Ciencias Aplicadas,Universidad Icesi, 2Centro de Investigaciones Microbiológicas (CIMIC), Department of Biological Sciences,Universidad de los Andes, 3BioDx: Diagnóstico y Soluciones Biotecnológicas S.A.S,Universidad Icesi

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Summary

在这里,我们提供了一个完整的协议,用于标准化和实施通过逆转录环介导的等温扩增 (RT-LAMP) 检测人类样本中 SARS-CoV-2 病毒的方法。这种方法在 60 分钟内完成,可以以低成本和廉价的设备适用于任何实验室或护理点。

Transcript

我们的研究旨在将 LAMP 标准化为诊断前测试,适用于缺乏先进实验室基础设施的现场或初级保健环境。我们寻求促进使用本地可生产的缓冲液和替代染料,减少对进口商业试剂盒的依赖。目前,CRISPR-Cas是一种高效的系统,用于开发快速、特异性和灵敏的病原体诊断方法。

现在,该系统具有广泛的补充范围,可检测不同目标样品中的 RNA 或 DNA 靶标。目前的实验挑战涉及防止由扩增子污染和原代细胞引起的非靶标扩增,特别是对于容易形成二聚体的长引物。此外,我们的目标是确定最佳反应条件,以实现染料中视觉上可区分的颜色变化。

一旦标准化并得到充分验证,该方法就可以在60分钟内轻松实施并适应任何实验室或护理点,以低成本和廉价的设备,检测几乎任何病原体。因此,它有可能在初级保健中心用于进行及时的流行病学监测。我们的多学科团队专注于从哥伦比亚本土植物中发现分子,试图确定抑制登革热、寨卡病毒或 SARS-COVID-2 等病毒复制的潜力。

此外,我们正在进行研究,以开发这些病毒的分子诊断方法。

Explore More Videos

关键词:SARS-CoV-2 逆转录 环介导等温扩增 (RT-LAMP) CRISPR-CAS 分子诊断 病原体检测 现场检测 即时护理 低成本 植物源性抑制剂

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