Pöhlmann Christopher, Wang Yiran, Humenik Martin, Heidenreich Bernd, Gareis Manfred, Sprinzl Mathias
Laboratorium für Biochemie, Universität Bayreuth, Universitätsstrasse 30, 95440 Bayreuth, Germany.
Biosens Bioelectron. 2009 May 15;24(9):2766-71. doi: 10.1016/j.bios.2009.01.042. Epub 2009 Feb 6.
Electrochemical biochips are an emerging tool for point-of-care diagnostic systems in medicine, food and environmental monitoring. In the current study, a thermostable reporter enzyme, esterase 2 (EST2) from Alicyclobacillus acidocaldarius, is used for specific and sensitive detection of bacteria by one-step rRNA/DNA hybridization between a bacterium-specific capture oligodeoxynucleotide (ODN), bacterial 16S rRNA and an uniform EST2-ODN reporter conjugate. The detection limit corresponds to approximately 500 colony forming units (cfu) Escherichia coli. Beside high sensitivity, the application of electrochemical biochips allows discrimination of two gram-negative and two gram-positive bacteria demonstrating the specificity and the potential for parallel detection of microorganisms. The feasibility of identification of foodborne bacteria was studied with meat juice contaminated with E. coli. This detection system has the capability to be applied for monitoring of bacterial food contamination.
电化学生物芯片是医学、食品和环境监测中即时诊断系统的一种新兴工具。在本研究中,一种来自嗜酸嗜热栖热菌的热稳定报告酶酯酶2(EST2),被用于通过细菌特异性捕获寡脱氧核苷酸(ODN)、细菌16S rRNA与均匀的EST2-ODN报告偶联物之间的一步rRNA/DNA杂交,对细菌进行特异性和灵敏的检测。检测限约为500个大肠杆菌菌落形成单位(cfu)。除了高灵敏度外,电化学生物芯片的应用还能够区分两种革兰氏阴性菌和两种革兰氏阳性菌,证明了其特异性以及平行检测微生物的潜力。利用被大肠杆菌污染的肉汁研究了食源细菌鉴定的可行性。该检测系统有能力应用于监测细菌性食品污染。