Klaschik S, Lehmann L E, Raadts A, Book M, Gebel J, Hoeft A, Stuber F
Department of Anesthesiology and Intensive Care Medicine. Institute for Hygiene and Public Health, University of Bonn, Bonn, Germany.
J Clin Microbiol. 2004 Feb;42(2):512-7. doi: 10.1128/JCM.42.2.512-517.2004.
We introduce a consensus real-time PCR protocol for the detection of bacterial DNA from laboratory-prepared specimens such as water, urine, and plasma. This prototype detection system enables an exact Gram stain classification and, in particular, screening for specific species of 17 intensive care unit-relevant bacteria by means of fluorescence hybridization probes and melting-curve analysis in a one-run experiment. One strain of every species was tested at a final density of 10(6) CFU/ml. All bacteria examined except Staphylococcus aureus and Staphylococcus epidermidis could be differentiated successfully; S. aureus and S. epidermidis could only be classified as "Staphylococcus species." The hands-on time for preparation of the DNA, performance of the PCR, and evaluation of the PCR results was less than 4 h. Nevertheless, this prototype detection system requires more clinical validation.
我们介绍了一种用于从实验室制备的样本（如水、尿液和血浆）中检测细菌DNA的实时聚合酶链反应（PCR）共识方案。这种原型检测系统能够进行精确的革兰氏染色分类，特别是通过荧光杂交探针和熔解曲线分析，在一次实验中筛选出17种与重症监护病房相关的特定细菌种类。每种细菌的一个菌株在最终密度为10(6) CFU/ml时进行了测试。除金黄色葡萄球菌和表皮葡萄球菌外，所有检测的细菌都能成功区分；金黄色葡萄球菌和表皮葡萄球菌只能归类为“葡萄球菌属”。DNA制备、PCR操作以及PCR结果评估的实际操作时间不到4小时。然而，这种原型检测系统需要更多的临床验证。
