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使用环介导等温扩增技术(LAMP)对空气传播病原体进行快速现场检测。

Rapid In-Field Detection of Airborne Pathogens Using Loop-Mediated Isothermal Amplification (LAMP).

作者信息

Bani Alessia, Whitby Corinne, Colbeck Ian, Dumbrell Alex J, Ferguson Robert M W

机构信息

School of Life Sciences, University of Essex, Wivenhoe Park, Colchester, CO4 3SQ, UK.

Aquatic Research Facility, Nature-Based Solutions Research Centre, University of Derby, Derby DE22 1GB, UK.

出版信息

Microorganisms. 2024 Dec 13;12(12):2578. doi: 10.3390/microorganisms12122578.

Abstract

Multiple human and plant pathogens are dispersed and transmitted as bioaerosols (e.g., , SARS-CoV-2, , , spp., and ). Rapid, on-site methods to detect airborne pathogens would greatly enhance our ability to monitor exposure and trigger early mitigation measures across different settings. Analysis of air samples for microorganisms in a regulatory context is often based on culture-based methods, which are slow, lack specificity, and are not suitable for detecting viruses. Molecular methods (based on nucleic acids) could overcome these challenges. For example, loop-mediated isothermal amplification (LAMP) is rapid, sensitive, specific, and may detect microbial pathogens from air samples in under 60 min. However, the low biomass in air samples makes recovering sufficient nucleic acids for detection challenging. To overcome this, we present a simple method for concentrating bioaerosols collected through liquid impingement (one of the most common methods for bioaerosol collection). This method paired with LAMP (or other molecular approaches) offers simple, rapid, and sensitive detection of pathogens. We validated this method using three airborne pathogens (, , and ), and we were able to detect fewer than five cells in a 15 mL liquid impinger air sample in under 60 min. This simple method offers rapid pathogen detection without the use of specialist equipment, and it can be used across healthcare, education, environmental monitoring, and military settings.

摘要

多种人类和植物病原体以生物气溶胶的形式传播(例如,严重急性呼吸综合征冠状病毒2、 、 、 属和 )。快速的现场空气传播病原体检测方法将大大提高我们在不同环境中监测暴露情况并触发早期缓解措施的能力。在监管环境下对空气样本中的微生物进行分析通常基于培养方法,这些方法速度慢、缺乏特异性,且不适用于检测病毒。分子方法(基于核酸)可以克服这些挑战。例如,环介导等温扩增(LAMP)快速、灵敏、特异,可在60分钟内从空气样本中检测出微生物病原体。然而,空气样本中的生物量较低,难以回收足够用于检测的核酸。为克服这一问题,我们提出了一种简单的方法,用于浓缩通过液体冲击法(生物气溶胶收集最常用的方法之一)收集的生物气溶胶。该方法与LAMP(或其他分子方法)相结合,可实现对病原体的简单、快速且灵敏的检测。我们使用三种空气传播病原体( 、 、 )对该方法进行了验证,能够在60分钟内从15 mL液体冲击器空气样本中检测到少于五个细胞。这种简单的方法无需使用专业设备即可快速检测病原体,可用于医疗保健、教育、环境监测和军事环境等领域。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aeab/11678261/7b224659014d/microorganisms-12-02578-g001.jpg

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