Gabig-Ciminska Magdalena, Los Marcin, Holmgren Anders, Albers Jörg, Czyz Agata, Hintsche Reiner, Wegrzyn Grzegorz, Enfors Sven Olof
Department of Biotechnology, Royal Institute of Technology (KTH), Roslagstullsbacken 21, S-10691 Stockholm, Sweden.
Anal Biochem. 2004 Jan 1;324(1):84-91. doi: 10.1016/j.ab.2003.09.020.
Infections of bacterial cultures by bacteriophages are common and serious problems in many biotechnological laboratories and factories. A method for specific, quantitative, and quick detection of phage contamination, based on the use of electric DNA chip is described here. Different phages of Escherichia coli and Bacillus subtilis were analyzed. Phage DNA was isolated from bacterial culture samples and detected by combination of bead-based sandwich hybridization with enzyme-labeled probes and detection of the enzymatic product using silicon chips. The assay resulted in specific signals from all four tested phages without significant background. Although high sensitivity was achieved in 4h assay time, a useful level of sensitivity (10(7)-10(8) phages) is achievable within 25 min. A multiplex DNA chip technique involving a mixture of probes allows for detection of various types of phages in one sample. These analyses confirmed the specificity of the assay.
在许多生物技术实验室和工厂中，噬菌体对细菌培养物的感染是常见且严重的问题。本文描述了一种基于使用电DNA芯片的特异性、定量且快速检测噬菌体污染的方法。对大肠杆菌和枯草芽孢杆菌的不同噬菌体进行了分析。从细菌培养物样品中分离出噬菌体DNA，并通过基于磁珠的夹心杂交与酶标记探针相结合以及使用硅芯片检测酶产物来进行检测。该检测方法在所有四种测试噬菌体中均产生了特异性信号，且背景不显著。虽然在4小时的检测时间内实现了高灵敏度，但在25分钟内也可达到有用的灵敏度水平（10⁷ - 10⁸个噬菌体）。涉及探针混合物的多重DNA芯片技术可在一个样品中检测多种类型的噬菌体。这些分析证实了该检测方法的特异性。
