Rapid detection of total and viable Legionella pneumophila in tap water by immunomagnetic separation, double fluorescent staining and flow cytometry.
Rapid detection of total and viable Legionella pneumophila in tap water by immunomagnetic separation, double fluorescent staining and flow cytometry.
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DOI：
10.1111/j.1751-7915.2012.00366.x
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发表时间：
2012-11
影响因子：
5.7
通讯作者：
Egli T
中科院分区：
文献类型：
--
作者：
Keserue HA;Baumgartner A;Felleisen R;Egli T
我们建立了一种快速检测嗜肺军团菌（Lp）的方法，通过过滤，免疫磁性分离，双重荧光染色，流式细胞术（IMS-FCM方法）。该方法需要120分钟，可以区分“活”和“膜损伤”细胞。回收率超过85%的加标Lp SG 1细胞在1升的自来水和检测限约为50和15个细胞每升的总和活的Lp，分别。在一项盲法研究中，使用来自家庭设施的水样对该方法进行了比较，其中三个环境实验室采用ISO 11731电镀法。在53%的水样品从不同的水龙头和淋浴显着较高浓度的脂蛋白检测流式细胞仪。未发现与平板培养方法相关。由于我们的方法也可检测到“活但不可培养”（VNBC）细胞，因此该结果符合预期。IMS-FCM方法受到所用抗体特异性的限制;在本例中，它们靶向Lp血清群1-12。这一点以及未检出含Lp阿米巴原虫的事实可能解释了为什么使用平板培养法在21%的所有样本中观察到较高的计数。虽然IMS‐FCM方法还不适合完全取代常规Lp监测的既定平板法（ISO 11731），但它具有平板法的主要优势，可以快速提供对配水系统中这种病原体生态学的重要见解。
We developed a rapid detection method for Legionella pneumophila (Lp) by filtration, immunomagnetic separation, double fluorescent staining, and flow cytometry (IMS‐FCM method). The method requires 120 min and can discriminate ‘viable’ and ‘membrane‐damaged’ cells. The recovery is over 85% of spiked Lp SG 1 cells in 1 l of tap water and detection limits are around 50 and 15 cells per litre for total and viable Lp, respectively. The method was compared using water samples from house installations in a blind study with three environmental laboratories performing the ISO 11731 plating method. In 53% of the water samples from different taps and showers significantly higher concentrations of Lp were detected by flow cytometry. No correlation to the plate culture method was found. Since also ‘viable but not culturable’ (VNBC) cells are detected by our method, this result was expected. The IMS‐FCM method is limited by the specificity of the used antibodies; in the presented case they target Lp serogroups 1–12. This and the fact that no Lp‐containing amoebae are detected may explain why in 21% of all samples higher counts were observed using the plate culture method. Though the IMS‐FCM method is not yet fit to completely displace the established plating method (ISO 11731) for routine Lp monitoring, it has major advantages to plating and can quickly provide important insights into the ecology of this pathogen in water distribution systems.
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影响因子：
12.8
作者：
Hammes, Frederik;Berney, Michael;Egli, Thomas
通讯作者：
Egli, Thomas
影响因子：
4.4
作者：
Allegra, Severine;Grattard, Florence;Berthelot, Philippe
通讯作者：
Berthelot, Philippe
影响因子：
2.6
作者：
Bentham, RH
通讯作者：
Bentham, RH
影响因子：
4.4
作者：
Allegra, Severine;Berger, Francoise;Riffard, Serge
通讯作者：
Riffard, Serge
影响因子：
4.4
作者：
Aurell, H;Catala, P;Lebaron, P
通讯作者：
Lebaron, P