集成空气采样和环介导等温扩增(LAMP)技术在室内环境中检测生物气溶胶的发展

A. Patange, Zhihang Zhang, Ruairi Monaghan, M. Fallon, H. Humphreys, B. Tiwari, S. Daniels
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引用次数: 0

摘要

接触生物气溶胶与一系列广泛的公共卫生问题有关。致病性生物气溶胶可导致各种疾病的发病,因此快速有效地检测它们对公共卫生至关重要。环介导等温扩增(LAMP)是一种特异性高、准确度高的核酸扩增方法。在这项研究中,我们开发了一种简化的LAMP测定方法,能够以最小的化学和加工要求检测气溶胶中的微生物。设计了一种空气采样系统,用于有效地收集和回收气溶胶中的微生物,并将其集成到LAMP分析过程中。我们展示了大肠杆菌(E. coli)气溶胶的成功收集和比色LAMP检测。结果发现,比色LAMP法检测到的大肠杆菌浓度低至102 CFU/ml。这种组合技术能够在常规实验室环境之外准确快速地进行生物气溶胶的基因组检测。这项工作描述了一个全自动比色LAMP测定装置,Luremain在室温下稳定长达4周,然而这项研究正在进行中,我们期望试剂的使用寿命显着延长。smAir LM365,便于集成技术,操作简便。将空气采样、DNA提取、DNA扩增、检测等过程集成在该装置上。在作业者要求进行干预之前,该装置可以完成多个检测过程。我们证明了大肠杆菌污染的水样可以在我们的LAMP检测设备上在大约60分钟内自动检测和分析。随着设备的自动化,LAMP试剂的稳定和长期储存是一个重要的要求。本文还对LAMP试剂的保存方法进行了评述,并对保存试剂在常温下的稳定性进行了评价。我们的数据表明,保存的LAMP试剂在室温下可以保持稳定长达4周,但这项研究仍在进行中,我们预计试剂的使用寿命会明显延长。
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Development of an Integrated Air Sampling and Loop-Mediated Isothermal Amplification (LAMP) technology for detection of bioaerosols in indoor environments
Exposure to bioaerosols are associated with wide a range of public health issues. Pathogenic bioaerosols can contribute to the onset of various diseases, therefore their rapid and efficient detection is crucial to public health. Loop Mediated Isothermal Amplification (LAMP) is a highly specific and accurate nucleic acid amplification method to detect microbes. In this study, we developed a simplified LAMP assay capable of detecting microbes in aerosols with minimal chemical and processing requirements. An air sampling system was designed to efficiently collect and recover microbes in aerosols and integrate into a LAMP assay process. We demonstrated successful collection of Escherichia coli (E. coli) aerosols and detection by a colorimetric LAMP assay. It was found that the colorimetric LAMP assay detected E. coli in concentrations as low as 102 CFU/ml. This combined technology enables accurate and rapid genomic detection of bioaerosols outside of conventional laboratory settings. This work describes a fully automated colorimetric LAMP assay device, the Luremain stable for up to 4 weeks at room temperature, however this study is ongoing, and we expect a significantly longer life of the reagent.smAir LM365, for facilitating the integrated technology with easy operation. All the processes including air sampling, DNA extraction, DNA amplification and detection were integrated on this device. The cartridge design allows the device to complete several detection processes before an intervention is required by an operator. We demonstrated that E. coli contaminated water samples can be automatically detected and analysed on our LAMP assay device in approximately 60 min. Along with the automation of the device, stable and long-term storage of LAMP reagents is an important requirement. Here we also comment on a preservation method for the LAMP reagents, and we evaluate the stability of preserved reagents at ambient temperature. Our data indicate that preserved LAMP reagents can remain stable for up to 4 weeks at room temperature, however this study is ongoing, and we expect a significantly longer life of the reagent.
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