Schafer Millie P, Martinez Kenneth F, Mathews Elaine S
National Institute for Occupational Safety and Health, Cincinnati, Ohio, USA.
Appl Occup Environ Hyg. 2003 Jan;18(1):41-50. doi: 10.1080/10473220301387.
Novel environmental air and water mycobacteria sampling and analytical methods are needed to circumvent difficulties associated with the use of culture-based methodologies. To implement this objective, a commercial, clinical, genus DNA amplification method utilizing the polymerase chain reaction (PCR) was interfaced with novel air sampling strategies in the laboratory. Two types of air samplers, a three-piece plastic, disposable filter cassette and an eight-stage micro-orifice uniform deposit impactor (MOUDI), were used in these studies. In both samplers, 37-mm polytetrafluoroethylene (PTFE) filters were used. Use of the MOUDI sampler permitted the capture of airborne mycobacteria in discrete size ranges, an important parameter for relating the airborne mycobacteria cells to potential respirable particles (aerodynamic diameter <10 microm) capable of causing health effects. Analysis of the samples was rapid, requiring only 1-1.5 days, as no microbial culturing or DNA purification was required. This approach was then used to detect suspected mycobacteria contamination associated with pools at a large public facility. PCR was also used to analyze various water samples from these pools. Again, no culturing or sample purification was required. Water samples taken from all ultraviolet light/hydrogen peroxide-treated whirlpools tested positive for the presence of mycobacteria. No mycobacteria were detected in the chlorine-treated pools and the water main supply facility. All air samples collected in the proximity of the indoor whirlpools and the associated changing rooms were strongly positive for airborne mycobacteria. The airborne mycobacteria particles were predominantly collected on MOUDI stages 1-6 representing an aerodynamic size range of 0.5 to 9.9 microm. In conclusion, using this approach permits the rapid detection of mycobacteria contamination as well as the routine monitoring of suspected pools. The approach circumvents problems associated with culture-based methods such as fungal overgrowth on agar plates, and the presence of nonculturable or difficult to culture mycobacteria strains.
需要新的环境空气和水的分枝杆菌采样及分析方法，以克服基于培养的方法所带来的困难。为实现这一目标，在实验室中，一种利用聚合酶链反应（PCR）的商业、临床、属DNA扩增方法与新的空气采样策略相结合。在这些研究中使用了两种类型的空气采样器，一种三件式塑料一次性滤膜盒和一个八级微孔均匀沉积冲击器（MOUDI）。在这两种采样器中，均使用了37毫米的聚四氟乙烯（PTFE）滤膜。使用MOUDI采样器能够捕获不同大小范围的空气中分枝杆菌，这是将空气中的分枝杆菌细胞与可能导致健康影响的潜在可吸入颗粒（空气动力学直径<10微米）相关联的一个重要参数。样品分析速度很快，仅需1 - 1.5天，因为无需进行微生物培养或DNA纯化。然后该方法被用于检测一个大型公共设施中与水池相关的疑似分枝杆菌污染。PCR还被用于分析这些水池的各种水样。同样，无需进行培养或样品纯化。从所有经紫外线/过氧化氢处理的漩涡池中采集的水样检测出分枝杆菌呈阳性。在经过氯处理的水池和主要供水设施中未检测到分枝杆菌。在室内漩涡池及其相关更衣室附近采集的所有空气样本中，空气中分枝杆菌均呈强阳性。空气中的分枝杆菌颗粒主要收集在MOUDI的第1 - 6级，代表空气动力学尺寸范围为0.5至9.9微米。总之，使用这种方法能够快速检测分枝杆菌污染，并对疑似水池进行常规监测。该方法避免了基于培养的方法所存在的问题，如琼脂平板上真菌过度生长，以及存在不可培养或难以培养的分枝杆菌菌株。
