Pyankov Oleg V, Agranovski Igor E, Pyankova Olga, Mokhonova Ekaterina, Mokhonov Vlad, Safatov Alexander S, Khromykh Alexander A
Faculty of Environmental Sciences, Griffith University, Brisbane, 4111, Qld, Australia.
Environ Microbiol. 2007 Apr;9(4):992-1000. doi: 10.1111/j.1462-2920.2006.01226.x.
We have recently developed a new personal sampler and demonstrated its feasibility for detection of viable airborne microorganisms including bacteria, fungi and viruses. To accelerate the time-consuming analytical procedure involving 2-5 days of biological testing, we employed a real-time PCR protocol in conjunction with the personal sampler for collection of airborne viruses. The advantage of this approach is that if the presence of a particular pathogen in the air is detected by the PCR, the remaining collecting liquid can be further analysed by more time-consuming biological methods to estimate the number of airborne infectious/live microorganisms. As sampling of bioaerosols in natural environments is likely to be associated with substantial contamination by a range of microorganisms commonly existing in an ambient air, an investigation of the specificity of detection by targeted PCR analysis is required. Here we present the results of the study on the detection of Influenza virus in the ambient air contaminated with high concentrations of bacteria and fungi using real-time PCR protocol. The combined sampling PCR detection method was found to be fully feasible for the rapid ( approximately 2.5 h) and highly specific (no cross-reactivity) identification of the labile airborne virus in the air containing elevated concentrations of other microorganisms.
我们最近开发了一种新型个人采样器,并证明了其在检测包括细菌、真菌和病毒在内的空气传播活微生物方面的可行性。为了加速耗时2至5天的生物检测分析过程,我们采用了实时PCR方案,并结合个人采样器来收集空气传播病毒。这种方法的优点是,如果通过PCR检测到空气中存在特定病原体,剩余的收集液可以通过更耗时的生物方法进一步分析,以估计空气传播的感染性/活微生物数量。由于在自然环境中采集生物气溶胶可能会受到环境空气中普遍存在的一系列微生物的大量污染,因此需要对靶向PCR分析的检测特异性进行研究。在此,我们展示了使用实时PCR方案检测高浓度细菌和真菌污染的环境空气中流感病毒的研究结果。结果发现,联合采样PCR检测方法对于快速(约2.5小时)和高度特异性(无交叉反应)鉴定含有高浓度其他微生物的空气中不稳定的空气传播病毒是完全可行的。