1Infectious Diseases Division,Central Texas Veterans Health Care System,Clinical Associate Professor Department of Medicine,Texas A&M Health Science Center,Bryan,Texas.
2Cooperative Studies Program Coordinating Center,VA Maryland Health Care System,Perry Point,Maryland.
Infect Control Hosp Epidemiol. 2018 Mar;39(3):259-266. doi: 10.1017/ice.2017.318. Epub 2018 Feb 5.
OBJECTIVES We conducted this study to determine what sample volume, concentration, and limit of detection (LOD) are adequate for environmental validation of Legionella control. We also sought to determine whether time required to obtain culture results can be reduced compared to spread-plate culture method. We also assessed whether polymerase chain reaction (PCR) and in-field total heterotrophic aerobic bacteria (THAB) counts are reliable indicators of Legionella in water samples from buildings. DESIGN Comparative Legionella screening and diagnostics study for environmental validation of a healthcare building water system. SETTING Veterans Health Administration (VHA) facility water system in central Texas. METHODS We analyzed 50 water samples (26 hot, 24 cold) from 40 sinks and 10 showers using spread-plate cultures (International Standards Organization [ISO] 11731) on samples shipped overnight to the analytical lab. In-field, on-site cultures were obtained using the PVT (Phigenics Validation Test) culture dipslide-format sampler. A PCR assay for genus-level Legionella was performed on every sample. RESULTS No practical differences regardless of sample volume filtered were observed. Larger sample volumes yielded more detections of Legionella. No statistically significant differences at the 1 colony-forming unit (CFU)/mL or 10 CFU/mL LOD were observed. Approximately 75% less time was required when cultures were started in the field. The PCR results provided an early warning, which was confirmed by spread-plate cultures. The THAB results did not correlate with Legionella status. CONCLUSIONS For environmental validation at this facility, we confirmed that (1) 100 mL sample volumes were adequate, (2) 10× concentrations were adequate, (3) 10 CFU/mL LOD was adequate, (4) in-field cultures reliably reduced time to get results by 75%, (5) PCR provided a reliable early warning, and (6) THAB was not predictive of Legionella results. Infect Control Hosp Epidemiol 2018;39:259-266.
目的 本研究旨在确定环境验证军团菌控制所需的样本量、浓度和检测限(LOD),并寻求与平板扩散培养法相比,能否缩短获得培养结果所需的时间。还评估了聚合酶链反应(PCR)和现场总异养需氧菌(THAB)计数是否可作为建筑物水样中军团菌的可靠指标。
设计 用于医疗建筑水系统环境验证的比较性军团菌筛选和诊断研究。
地点 德克萨斯州中部退伍军人事务部(VHA)设施水系统。
方法 对来自 40 个水槽和 10 个淋浴器的 50 个水样(26 个热水,24 个冷水)进行分析,使用过夜运至分析实验室的平板扩散培养法(国际标准组织[ISO] 11731)对样品进行分析。现场采用 PVT(Phigenics Validation Test)培养滴片式采样器进行现场培养。对每个样品进行属级军团菌 PCR 检测。
结果 无论过滤的样本量如何,均未观察到明显差异。较大的样本量可检测到更多的军团菌。在 1 个菌落形成单位(CFU)/mL 或 10 CFU/mL LOD 时,未观察到统计学上的显著差异。当在现场开始培养时,所需时间可减少约 75%。PCR 结果提供了早期预警,经平板扩散培养得到证实。THAB 结果与军团菌状态无关。
结论 对于该设施的环境验证,我们确认(1)100mL 样本量足够,(2)10 倍浓度足够,(3)10 CFU/mL LOD 足够,(4)现场培养可将获得结果的时间可靠地缩短 75%,(5)PCR 提供了可靠的早期预警,(6)THAB 不能预测军团菌结果。
感染控制与医院流行病学 2018;39:259-266.