Department of Chemistry and Biochemistry, University of Texas at Arlington, Arlington, TX 76019, USA.
J Pharm Biomed Anal. 2010 Sep 21;53(1):75-80. doi: 10.1016/j.jpba.2010.03.010. Epub 2010 Mar 16.
Detection of microbial contamination in blood plasma is critical and necessary in different medical and research fields. Most of the current standard procedures for the detection of bacteria and fungi can be time-consuming, for example, direct inoculation methods of microbial cultures in respective growth media can take a few days to several weeks. A fast analysis method with high sensitivity output such as CE-laser-induced fluorescence becomes an attractive alternative. Previously, a spacer-injection method with the use of zwitterionic surfactant (SB3-10) as a blocking agent to negate the cells' mobility, induce aggregation and single microbial peak formation in a buffer solution was reported. Here, a fast, simple direct method for microbial detection in blood plasma without using the spacer and blocking agent is reported. To compensate for the natural electrophoretic heterogeneity of microbes, a CTAB additive was used to sweep all microbial cells towards the plasma peak where a single sharp microbial peak is formed and detected. With the use of BacLight Green bacterial stain, the microbial peak, generally, can be detected within 10 min in front of the plasma peak using capillary electrophoresis coupled with laser-induced fluorescence detection. The LOD of microbes detectable were 5 cells per injection. This technique provides a great advantage over traditional, time-consuming microbial inoculation methods.
检测血液血浆中的微生物污染在不同的医学和研究领域都是至关重要且必要的。目前大多数用于检测细菌和真菌的标准程序都很耗时,例如,将微生物培养物直接接种到各自的生长培养基中的方法可能需要几天到几周的时间。具有高灵敏度输出的快速分析方法,如 CE-激光诱导荧光,成为一种有吸引力的替代方法。之前,已经报道了一种使用两性离子表面活性剂(SB3-10)作为阻断剂的间隔注入方法,以消除细胞的迁移性,在缓冲溶液中诱导聚集和单个微生物峰形成。在此,报道了一种无需使用间隔物和阻断剂即可在血液血浆中快速、简单地直接检测微生物的方法。为了补偿微生物的天然电泳异质性,使用 CTAB 添加剂将所有微生物细胞扫向等离子体峰,在那里形成并检测到单个尖锐的微生物峰。使用 BacLight Green 细菌染色剂,使用毛细管电泳结合激光诱导荧光检测,通常可以在血浆峰前 10 分钟内检测到微生物峰。可检测到的微生物的 LOD 为 5 个细胞/注射。与传统的耗时的微生物接种方法相比,该技术具有很大的优势。