Environmental Microbiology Unit, Department of Public Health, School of Medicine, University of Patras, Rion, GR 26504, Greece.
Virol J. 2011 Jul 27;8:369. doi: 10.1186/1743-422X-8-369.
Airborne viruses remain one of the major public health issues worldwide. Detection and quantification of airborne viruses is essential in order to provide information regarding public health risk assessment.
In this study, an optimized new, simple, low cost method for sampling of airborne viruses using Low Melting Agarose (LMA) plates and a conventional microbial air sampling device has been developed. The use of LMA plates permits the direct nucleic acids extraction of the captured viruses without the need of any preliminary elution step. Molecular detection and quantification of airborne viruses is performed using real-time quantitative (RT-)PCR (Q(RT-)PCR) technique. The method has been tested using Adenoviruses (AdVs) and Noroviruses (NoVs) GII, as representative DNA and RNA viruses, respectively. Moreover, the method has been tested successfully in outdoor experiments, by detecting and quantifying human adenoviruses (HAdVs) in the airborne environment of a wastewater treatment plant.
The great advantage of LMA is that nucleic acids extraction is performed directly on the LMA plates, while the eluted nucleic acids are totally free of inhibitory substances. Coupled with QPCR the whole procedure can be completed in less than three (3) hours.
空气中的病毒仍然是全球主要的公共卫生问题之一。为了提供有关公共卫生风险评估的信息，检测和定量空气中的病毒至关重要。
在这项研究中，开发了一种使用低熔点琼脂糖（LMA）平板和传统微生物空气采样设备的优化的新型、简单、低成本的空气病毒采样方法。使用 LMA 平板可以直接提取捕获病毒的核酸，而无需任何初步洗脱步骤。使用实时定量（RT-）PCR（Q（RT-）PCR）技术进行空气病毒的分子检测和定量。该方法已分别使用腺病毒（AdVs）和诺如病毒（NoVs）GII 作为代表性的 DNA 和 RNA 病毒进行了测试。此外，该方法已在户外实验中成功测试，通过在污水处理厂的空气环境中检测和定量人腺病毒（HAdVs）。
LMA 的巨大优势在于核酸直接在 LMA 平板上提取，而洗脱的核酸完全不含抑制物质。与 QPCR 结合，整个过程可以在三个（3）小时内完成。
