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首页> 外文期刊>Journal of Clinical Microbiology >Rapid Diagnosis of Bacteremia by Universal Amplification of 23S Ribosomal DNA Followed by Hybridization to an Oligonucleotide Array
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Rapid Diagnosis of Bacteremia by Universal Amplification of 23S Ribosomal DNA Followed by Hybridization to an Oligonucleotide Array

机译：通过普遍扩增23S核糖体DNA，然后杂交至寡核苷酸阵列，快速诊断细菌血症。

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摘要

The rapid identification of bacteria in blood cultures and other clinical specimens is important for patient management and antimicrobial therapy. We describe a rapid (<4 h) detection and identification system that uses universal PCR primers to amplify a variable region of bacterial 23S ribosomal DNA, followed by reverse hybridization of the products to a panel of oligonucleotides. This procedure was successful in discriminating a range of bacteria in pure cultures. When this procedure was applied directly to 158 unselected positive blood culture broths on the day when growth was detected, 125 (79.7%) were correctly identified, including 4 with mixed cultures. Nine (7.2%) yielded bacteria for which no oligonucleotide targets were present in the oligonucleotide panel, and 16 culture-positive broths (10.3%) produced no PCR product. In seven of the remaining eight broths, streptococci were identified but not subsequently grown, and one isolate of Staphylococcus aureus was misidentified as a coagulase-negative staphylococcus. The accuracy, range, and discriminatory power of the assay can be continually extended by adding further oligonucleotides to the panel without significantly increasing complexity or cost.
机译：快速鉴定血液培养物和其他临床标本中的细菌对于患者管理和抗菌治疗很重要。我们描述了使用通用PCR引物扩增细菌23S核糖体DNA可变区的快速（<4 h）检测和鉴定系统，然后将产物反向杂交到一组寡核苷酸上。此程序成功地区分了纯培养物中的多种细菌。当在检测到生长的当天将这种方法直接应用于158个未选择的阳性血液培养肉汤时，可以正确鉴定出125个（79.7％），包括4个混合培养。 9个（7.2％）产生的细菌在寡核苷酸组中不存在寡核苷酸靶标，并且16个培养阳性肉汤（10.3％）不产生PCR产物。在其余的八个肉汤中的七个中，鉴定出了链球菌，但随后没有生长，并且金黄色葡萄球菌的一种分离物被误认为是凝固酶阴性葡萄球菌。通过在面板上添加其他寡核苷酸，可以不断扩展测定的准确性，范围和区分能力，而不会显着增加复杂性或成本。

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