冻干逆转录酶和聚合酶用于SARS-CoV-2的即时诊断

IF 5.2 2区 生物学
Natalie Mutter, Barbara Posch, Yasmin Gillitschka, Nadezhda Kataeva, Birgit Michalek, Daniel Lager, Filip Staniszewski, Alexandra Schilder, Lidiya Osinkina, Maximilian Westenthanner, Joachim Stehr, Federico Buersgens, Johannes R. Peham
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引用次数: 0

摘要

早期诊断病原体是减少其传播和预防严重健康风险的关键。COVID-19大流行向我们展示了快速即时检测的必要性。在这里,我们描述了用于即时诊断的扩增主混合物的制备。为此,设计、表达和纯化了两种非专利扩增酶。关键部件的准备使其能够独立于交付问题、制造商组合和产品信息。为了长期储存和无冷运输,我们制作的扩增混合物被冻干。最后,我们将冻干母液应用于集成的即时诊断系统,可以检测到10拷贝/μL的COVID-19 RNA。稳定的无冷试剂与移动和低成本设备的组合将允许在现场或家庭环境中对病原体进行分子诊断。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Lyophilised Reverse Transcriptase and Polymerase for Point-of-Care Diagnostics of SARS-CoV-2

Lyophilised Reverse Transcriptase and Polymerase for Point-of-Care Diagnostics of SARS-CoV-2

Lyophilised Reverse Transcriptase and Polymerase for Point-of-Care Diagnostics of SARS-CoV-2

Lyophilised Reverse Transcriptase and Polymerase for Point-of-Care Diagnostics of SARS-CoV-2

Lyophilised Reverse Transcriptase and Polymerase for Point-of-Care Diagnostics of SARS-CoV-2

Lyophilised Reverse Transcriptase and Polymerase for Point-of-Care Diagnostics of SARS-CoV-2

Early diagnosis of pathogens is key to reducing their spreading and to preventing severe health risks. The COVID-19 pandemic showed us the need for rapid point-of-care tests. Here, we describe the preparation of an amplification master mix for point-of-care diagnostics. Therefore, two off-patent amplification enzymes were designed, expressed and purified. The preparation of the key components enables independence from delivery issues, manufacturer portfolio and product information's. For long-term storage and cold-free transport, our fabricated amplification mix was lyophilised. Finally, we applied our lyophilised master mix on an integrated point-of-care diagnostic system and could detect 10 copies/μL COVID-19 RNA. The combination of stable, cold-free reagents with the mobile and low-cost device will allow molecular diagnostics of pathogens in a field or home setting.

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来源期刊
Microbial Biotechnology
Microbial Biotechnology Immunology and Microbiology-Applied Microbiology and Biotechnology
CiteScore
11.20
自引率
3.50%
发文量
162
审稿时长
1 months
期刊介绍: Microbial Biotechnology publishes papers of original research reporting significant advances in any aspect of microbial applications, including, but not limited to biotechnologies related to: Green chemistry; Primary metabolites; Food, beverages and supplements; Secondary metabolites and natural products; Pharmaceuticals; Diagnostics; Agriculture; Bioenergy; Biomining, including oil recovery and processing; Bioremediation; Biopolymers, biomaterials; Bionanotechnology; Biosurfactants and bioemulsifiers; Compatible solutes and bioprotectants; Biosensors, monitoring systems, quantitative microbial risk assessment; Technology development; Protein engineering; Functional genomics; Metabolic engineering; Metabolic design; Systems analysis, modelling; Process engineering; Biologically-based analytical methods; Microbially-based strategies in public health; Microbially-based strategies to influence global processes
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