Subirats Jèssica, Royo Elena, Balcázar José Luis, Borrego Carles M
Catalan Institute for Water Research (ICRA), Scientific and Technological Park of the University of Girona, Girona, Spain.
Group of Molecular Microbial Ecology, Institute of Aquatic Ecology, University of Girona, Girona, Spain.
Environ Sci Pollut Res Int. 2017 Mar;24(7):6710-6714. doi: 10.1007/s11356-017-8426-6. Epub 2017 Jan 13.
In this study, we have developed real-time PCR assays using SYBR Green chemistry to detect all known alleles of bla , bla , and bla -like carbapenemase genes in water, sediment, and biofilm samples collected from hospital and wastewater treatment plant (WWTP) effluents and rivers receiving chronic WWTP discharges. The amplification of bla , bla , and bla DNA was linear over 7 log dilutions (R between 0.995 and 0.997) and showing efficiencies ranging from 92.6% to 100.3%. The analytical sensitivity indicated that the reaction for bla , bla , and bla -like genes was able to detect 35, 16, and 19 copy numbers per assay, respectively. The three carbapenemase genes were detected in hospital effluents, whereas only the bla and bla genes were detected in biofilm and sediment samples collected from wastewater-impacted rivers. The detection of bla , bla , and bla -like genes in different matrices suggests that carbapenem-resistant bacteria occur in both planktonic and benthic habitats thus expanding the range of resistance reservoirs for last-resort antibiotics. We believe that these real-time PCR assays would be a powerful tool for the rapid detection and quantification of bla , bla , and bla -like genes in complex environmental samples.
在本研究中，我们开发了使用SYBR Green化学的实时PCR检测方法，以检测从医院和污水处理厂（WWTP）废水以及接收长期WWTP排放的河流中采集的水、沉积物和生物膜样本中所有已知的bla、bla和bla样碳青霉烯酶基因等位基因。bla、bla和bla DNA的扩增在7个对数稀释度范围内呈线性（R在0.995至0.997之间），效率范围为92.6%至100.3%。分析灵敏度表明，bla、bla和bla样基因的反应每次检测分别能够检测到35、16和19个拷贝数。在医院废水中检测到了这三种碳青霉烯酶基因，而在从受废水影响的河流中采集的生物膜和沉积物样本中仅检测到了bla和bla基因。在不同基质中检测到bla、bla和bla样基因表明，耐碳青霉烯类细菌存在于浮游和底栖生境中，从而扩大了作为最后手段抗生素的耐药库范围。我们认为，这些实时PCR检测方法将成为快速检测和定量复杂环境样本中bla、bla和bla样基因的有力工具。
