The indiscriminate administration of broad-spectrum antibiotics is a primary contributor to the increasing prevalence of antibiotic resistance. Unfortunately, culture, the gold standard for bacterial identification is a time intensive process. Due to this extended diagnostic period, broad-spectrum antibiotics are generally prescribed to prevent poor outcomes. To overcome the deficits of culture-based methods, we have developed a rapid universal bacterial identification system. The platform utilizes a unique universal polymerase chain reaction (PCR) primer set that targets the internal transcribed spacer (ITS) regions between conserved bacterial genes, creating a distinguishable amplicon signature for every bacterial species. Bioinformatic simulation demonstrates that at least 45 commonly isolated pathogenic species can be uniquely identified using this approach. We experimentally confirmed these predictions on a representative set of pathogenic bacterial species. We also show that the system can determine the corresponding concentration of each pathogen. Finally, we validated performance in clinical urinary tract infection samples.

中文翻译：

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使用内部转录的间隔区靶向引物开发快速、免培养、通用的微生物鉴定系统


不加选择地使用广谱抗生素是抗生素耐药性日益普遍的主要原因。不幸的是，细菌鉴定的金标准培养是一个耗时的过程。由于诊断期延长，通常会开具广谱抗生素以防止不良结局。为了克服基于培养的方法的缺陷，我们开发了一种快速通用的细菌鉴定系统。该平台利用独特的通用聚合酶链式反应 （PCR） 引物组，靶向保守细菌基因之间的内部转录间隔区 （ITS） 区域，为每种细菌物种创建可区分的扩增子特征。生物信息学模拟表明，使用这种方法可以唯一识别至少 45 种常见分离的致病物种。我们在一组具有代表性的病原菌种类上实验证实了这些预测。我们还表明，该系统可以确定每种病原体的相应浓度。最后，我们在临床尿路感染样本中验证了性能。