Horká Marie, Růzicka Filip, Kubesová Anna, Holá Veronika, Slais Karel
Institute of Analytical Chemistry Academy of Sciences of the Czech Republic, 602 00 Brno, Czech Republic.
Anal Chem. 2009 May 15;81(10):3997-4004. doi: 10.1021/ac900374v.
In immunocompromised people fungal agents are able to cause serious infections with high mortality rate. An early diagnosis can increase the chances of survival of the affected patients. Simultaneously, the fungi produce toxins and they are frequent cause of allergy. Currently, various methods are used for detection and identification of these pathogens. They use microscopic examination and growth characteristic of the fungi. New methods are based on the analysis of structural elements of the target microorganisms such as proteins, polysaccharides, glycoproteins, nucleic acids, etc. for the construction of antibodies, probes, and primers for detection. The above-mentioned methods are time-consuming and elaborate. Here hydrophobic conidia from the cultures of different strains of the filamentous fungi were focused and separated by capillary zone electrophoresis and capillary isoelectric focusing. The detection was optimized by dynamic modifying of conidia by the nonionogenic tenside on the basis of pyrenebutanoate. Down to 10 labeled conidia of the fungal strains were fluorometrically detected, and isoelectric points of conidia were determined. The observed isoelectric points were compared with those obtained from the separation of the cultured clinical samples, and they were found to be not host-specific.
在免疫功能低下的人群中，真菌病原体能够引发死亡率很高的严重感染。早期诊断可以增加受感染患者的存活几率。同时，真菌会产生毒素，也是过敏的常见诱因。目前，有多种方法用于检测和鉴定这些病原体。这些方法利用真菌的显微镜检查和生长特性。新方法基于对目标微生物的结构成分（如蛋白质、多糖、糖蛋白、核酸等）进行分析，以构建用于检测的抗体、探针和引物。上述方法既耗时又复杂。在此，丝状真菌不同菌株培养物中的疏水性分生孢子通过毛细管区带电泳和毛细管等电聚焦进行聚焦和分离。基于芘丁酸的非离子表面活性剂对分生孢子进行动态修饰，从而优化了检测。荧光法检测到低至10个标记的真菌菌株分生孢子，并测定了分生孢子的等电点。将观察到的等电点与从培养的临床样本分离中获得的等电点进行比较，发现它们并非宿主特异性的。
