Pan Hongmiao, Zhang Yongbin, He Gui-Xin, Katagori Namrata, Chen Huizhong
Division of Microbiology, National Center for Toxicological Research, U,S, Food and Drug Administration, Jefferson 72079, AR, USA.
BMC Microbiol. 2014 Aug 21;14:222. doi: 10.1186/s12866-014-0222-6.
Due to potential interference of nanoparticles on bacterial quantification, there is a challenge to develop a fast, accurate and reproducible method for bacterial quantification. Currently various bacterial quantification methods are used by researchers performing nanoparticles study, but there has been no efficacy evaluation of these methods. Here we study interference of nanoparticles on three most commonly used conventional bacterial quantification methods, including colony counting to determine the colony-forming units (CFU), spectrophotometer method of optical density (OD) measurement, and flow cytometry (FCM).
Three oxide nanoparticles including ZnO, TiO2, and SiO2 and four bacterial species including Salmonella enterica serovar Newport, Staphylococcus epidermidis, Enterococcus faecalis, and Escherichia coli were included in the test. Results showed that there is no apparent interference of the oxide nanoparticles on quantifications of all four bacterial species by FCM measurement; CFU counting is time consuming, less accurate and not suitable for automation; and the spectrophotometer method using OD measurement was the most unreliable method to quantify and detect the bacteria in the presence of the nanoparticles.
In summary, FCM measurement proved to be the best method, which is suitable for rapid, accurate and automatic detection of bacteria in the presence of the nanoparticles.
由于纳米颗粒对细菌定量存在潜在干扰,因此开发一种快速、准确且可重复的细菌定量方法具有挑战性。目前,从事纳米颗粒研究的人员使用了各种细菌定量方法,但尚未对这些方法进行效能评估。在此,我们研究纳米颗粒对三种最常用的传统细菌定量方法的干扰,包括通过菌落计数来确定菌落形成单位(CFU)、利用分光光度计测量光密度(OD)的方法以及流式细胞术(FCM)。
测试中包括三种氧化物纳米颗粒,即氧化锌(ZnO)、二氧化钛(TiO2)和二氧化硅(SiO2),以及四种细菌,即肠炎沙门氏菌新港血清型(Salmonella enterica serovar Newport)、表皮葡萄球菌(Staphylococcus epidermidis)、粪肠球菌(Enterococcus faecalis)和大肠杆菌(Escherichia coli)。结果表明,氧化物纳米颗粒对通过流式细胞术测量的所有四种细菌的定量没有明显干扰;菌落形成单位计数耗时、准确性较低且不适用于自动化;在存在纳米颗粒的情况下,使用光密度测量的分光光度计方法是定量和检测细菌最不可靠的方法。
总之,流式细胞术测量被证明是最佳方法,适用于在存在纳米颗粒的情况下快速、准确且自动地检测细菌。