The rapid identification of pathogenic bacteria is important for determining an appropriate antimicrobial therapy for pneumonia, but traditional bacterial culture is time-consuming and labourious. The aim of this study was to develop and evaluate a DNA microarray assay for the simultaneous detection of fifteen bacterial species directly from respiratory tract specimens in patients with pneumonia. These species included Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus influenzae, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, Mycoplasma pneumoniae, Enterococcus faecalis, Enterococcus faecium, Enterobacter cloacae, Stenotrophomonas maltophilia, Burkholderia cepacia, Legionella pneumophila and Chlamydia pneumoniae. The 16S rDNA genes and other specific genes of each pathogen were chosen as the amplification targets, amplified via multiplex polymerase chain reaction (PCR), and hybridized to oligonucleotide probes in a microarray. The DNA microarray detection limit was 103 copies/μL. Nineteen standard strains and 119 clinical isolates were correctly detected with our microarray, and 3 nontarget species from 4 clinical isolates were not detected. Additionally, bacterial pathogens were accurately identified when two or three bacterial targets were mixed together. Furthermore, the results for 99.4% (156/157) of clinical specimens were the same as those from a conventional assay. We developed a DNA microarray that could simultaneously detect various bacterial pathogens in pneumonia. The method described here has the potential to provide considerable labour and time savings due to its ability to screen for 15 bacterial pathogens simultaneously.

中文翻译：

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用于快速检测肺炎中15种细菌病原体的DNA芯片检测方法的开发。

快速确定病原细菌对于确定适当的肺炎抗菌治疗很重要，但是传统的细菌培养既费时又费力。这项研究的目的是开发和评估可直接从呼吸道标本中同时检测肺炎患者中的15种细菌的DNA芯片检测方法。这些种类包括肺炎链球菌，金黄色葡萄球菌，流感嗜血杆菌，大肠杆菌，肺炎克雷伯菌，铜绿假单胞菌，鲍曼不动杆菌，支原体肺炎支原体，粪肠球菌，肺炎克雷伯氏菌，嗜碱杆菌，嗜酸性支原体，肠球菌，疟原虫，嗜中性肠炎球菌，疟原虫 选择每种病原体的16S rDNA基因和其他特异性基因作为扩增目标，通过多重聚合酶链反应（PCR）进行扩增，并与微阵列中的寡核苷酸探针杂交。DNA芯片检测限为103拷贝/μL。用我们的微阵列正确检测了19种标准菌株和119种临床分离株，并且未检测到4种临床分离株中的3种非靶标物种。另外，当将两个或三个细菌靶标混合在一起时，可以准确识别细菌病原体。此外，99.4％（156/157）的临床标本结果与常规测定的结果相同。我们开发了一种可同时检测肺炎中各种细菌病原体的DNA微阵列。