Bonetta Sara, Pignata Cristina, Bonetta Silvia, Meucci Lorenza, Giacosa Donatella, Marino Elena, Gilli Giorgio, Carraro Elisabetta
Department of Public Health and Pediatrics, University of Torino, P.zza Polonia 94, 10126 Torino, Italy.
Società Metropolitana Acque Torino S.p.A., C.so XI Febbraio, 14, 10152 Torino, Italy.
Int J Environ Res Public Health. 2017 Apr 27;14(5):467. doi: 10.3390/ijerph14050467.
is a ubiquitous microorganism widely distributed in aquatic environments and can cause Legionellosis in humans. A promising approach to detect viable cells in water samples involves the use of quantitative polymerase chain reaction (qPCR) in combination with photoactivatable DNA intercalator propidium monoazide (PMA). However, the PMA efficiency could be different depending on the experimental conditions used. The aim of this study was to compare two PMA exposure protocols: (A) directly on the membrane filter or (B) in liquid after filter washing. The overall PMA-induced qPCR means reductions in heat-killed cells were 2.42 and 1.91 log units for exposure protocols A and B, respectively. A comparison between the results obtained reveals that filter exposure allows a higher PMA-qPCR signal reduction to be reached, mainly at low concentrations ( < 0.05). This confirms the potential use of this method to quantify in water with low contamination.
是一种广泛存在于水生环境中的微生物,可导致人类感染军团病。一种检测水样中活细胞的有前景的方法是将定量聚合酶链反应(qPCR)与光活化DNA嵌入剂单叠氮溴化丙锭(PMA)结合使用。然而,PMA的效率可能因所使用的实验条件而异。本研究的目的是比较两种PMA暴露方案:(A)直接在膜滤器上或(B)在滤器洗涤后的液体中。对于暴露方案A和B,PMA诱导的qPCR对热灭活细胞的总体平均减少量分别为2.42和1.91个对数单位。对所得结果的比较表明,滤器暴露能够实现更高的PMA-qPCR信号降低,主要是在低浓度(<0.05)时。这证实了该方法在低污染水中定量[该微生物名称未给出]的潜在用途。