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Evaluation of Legionella presence in the water system of a public building by semi-nested polymerase chain reaction as a rapid screening method complementary to plate count

机译:通过半巢式聚合酶链反应评估军团菌在公共建筑水系统中的存在,作为平板计数的快速筛选方法

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The aim of this study was to apply a fast and efficient protocol for monitoring levels of Legionella contamination in high-risk water systems using molecular techniques and culture methods. Forty-nine water samples from a public building were analyzed by a culture method (BCYE agar) and a specific semi-nested polymerase chain reaction (PCR). The first 30 analyses using plate counts were positive in 28% of hot water and 8% of cold water samples. The analysis by PCR, obtained within 24 h of collection, revealed the presence of DNA in 100 and 54% respectively. Only four PCR results coincided with the cultures. A second survey (19 samples) was performed 1 year later. Semi-nested PCR revealed that 53% of the samples were positive; however, plate counts yielded no positive results. The 16S rRNA sequence comparison between the first and second samplings showed 100% homology. In conclusion, the study of the design of the building's water system, the use of a fast screening semi-nested PCR and a culture method for the detection of Legionella allowed accurate assessment of the contamination, thus contributing to the early implementation of measures to eliminate the presence of the bacteria in water systems and consequently reduce a latent public health risk.
机译:这项研究的目的是应用一种快速有效的方案,使用分子技术和培养方法监测高风险水系统中军团菌的污染水平。通过培养方法(BCYE琼脂)和特定的半巢式聚合酶链反应(PCR)分析了来自公共建筑的49个水样。使用平板计数的前30次分析在28%的热水和8%的冷水样品中均为阳性。收集后24小时内通过PCR进行的分析表明,分别有100%和54%的DNA存在。仅四个PCR结果与培养物一致。一年后进行了第二次调查(19个样本)。半巢式PCR显示53%的样本为阳性;但是,板数没有产生积极的结果。第一次和第二次采样之间的16S rRNA序列比较显示出100%的同源性。总之,对建筑物供水系统设计的研究,使用快速筛选半巢式PCR和培养方法检测军团菌可以准确评估污染,从而有助于尽早实施消除污染的措施水系统中细菌的存在,因此降低了潜在的公共健康风险。

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