Horká Marie, Tesařová Marie, Karásek Pavel, Růžička Filip, Holá Veronika, Sittová Martina, Roth Michal
Institute of Analytical Chemistry of the ASCR, v.v.i., Veveří 97, 602 00 Brno, Czech Republic.
Institute of Analytical Chemistry of the ASCR, v.v.i., Veveří 97, 602 00 Brno, Czech Republic.
Anal Chim Acta. 2015 Apr 8;868:67-72. doi: 10.1016/j.aca.2015.02.001. Epub 2015 Feb 4.
Serious bloodstream infections are a significant complication in critically ill patients. The treatment of these infections has become more difficult because of the increasing prevalence of multiresistant strains, especially methicillin-resistant Staphylococcus aureus (MRSA). Rapid differentiation of low number of MRSA from methicillin-susceptible S. aureus (MSSA) cells (10(1)-10(2) cells mL(-1)) in blood is necessary for fast effective antibiotic therapy. Currently, three groups of techniques, phenotyping, genotyping, and mass spectrometry, are used for MRSA and MSSA strains differentiation. Most of these techniques are time-consuming. PCR and other molecular techniques allow the detection and differentiation between MSSA and MRSA directly from blood cultures. These methods alone are rapid and they have good reproducibility and repeatability. Potential disadvantages of the genotyping methods include their discrimination ability, technical complexity, financial costs, and difficult interpretation of the results. Recently, capillary electrophoresis (CZE) was successfully used to differentiate between the agar-cultivated MRSA and MSSA strains in fused silica capillaries etched with supercritical water and modified with (3-glycidyloxypropyl)trimethoxysilane. The possible use of CZE as a fast and low-cost method for distinguishing between the blood-incubated MRSA or MSSA cells has been tested in this manuscript. Our goal was to test low amounts of bacteria (∼10(2) cell mL(-1)) similar to those in clinical samples. The migration times of the purified blood-incubated cells and the agar-cultivated cells were different from each other. However, their isoelectric point was the same for all strains.
严重的血流感染是重症患者的一种重要并发症。由于多重耐药菌株，尤其是耐甲氧西林金黄色葡萄球菌（MRSA）的患病率不断上升，这些感染的治疗变得更加困难。为了进行快速有效的抗生素治疗，有必要快速区分血液中少量的MRSA与甲氧西林敏感金黄色葡萄球菌（MSSA）细胞（10¹ - 10² 个细胞/mL）。目前，有三组技术，即表型分析、基因分型和质谱分析，用于区分MRSA和MSSA菌株。这些技术大多耗时较长。聚合酶链反应（PCR）和其他分子技术可直接从血培养物中检测并区分MSSA和MRSA。这些方法本身快速，且具有良好的重现性和重复性。基因分型方法的潜在缺点包括其鉴别能力、技术复杂性、财务成本以及结果解释困难。最近，毛细管电泳（CZE）已成功用于在经超临界水蚀刻并用（3 - 环氧丙氧基丙基）三甲氧基硅烷修饰的熔融石英毛细管中区分琼脂培养的MRSA和MSSA菌株。本文测试了CZE作为一种快速且低成本的方法来区分血液培养的MRSA或MSSA细胞的可能性。我们的目标是测试与临床样本中相似的少量细菌（约10² 个细胞/mL）。纯化的血液培养细胞和琼脂培养细胞的迁移时间彼此不同。然而，所有菌株的等电点相同。
