Automated Broad-Range Molecular Detection of Bacteria in Clinical Samples
Automated Broad-Range Molecular Detection of Bacteria in Clinical Samples
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DOI：
10.1128/jcm.02886-15
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发表时间：
2016-04-01
影响因子：
9.4
通讯作者：
Savelkoul, Paul H. M.
中科院分区：
文献类型：
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作者：
Budding, Andries E.;Hoogewerf, Martine;Savelkoul, Paul H. M.
分子检测方法，如定量PCR (qPCR)，已经进入临床微生物学实验室，用于检测一系列病原体。然而，大多数常规使用的方法都是针对特定物种的。因此，未显式搜索的任何内容都将丢失。这极大地限制了这些技术的灵活性和普遍应用。我们研究了一种快速通用细菌分子鉴定方法IS-pro在临床微生物实验室收到的常规患者样本中的应用。is -pro是一种基于16S-23S rRNA基因间隔区域的检测和分类的真细菌技术，其长度对每个微生物物种都是特异性的。由于这是一种开放的技术，临床医生不需要事先决定要寻找什么。我们使用66个样本进行常规细菌诊断测试，将常规培养与IS-pro进行比较。样本是从通常无菌部位（没有常驻微生物群）感染的患者中获得的。20份（30%）样品结果相同，31份（47%）样品IS-pro检测到的细菌种类多于培养样品，10份培养阴性样品中有5份IS-pro阳性。从这些培养阴性/IS-pro阳性样本中获得的5例患者的病例史表明，IS-pro的发现与临床高度相关。我们的研究结果表明，一种开放的分子方法，如IS-pro，可能对临床实践具有很高的附加价值。
Molecular detection methods, such as quantitative PCR (qPCR), have found their way into clinical microbiology laboratories for the detection of an array of pathogens. Most routinely used methods, however, are directed at specific species. Thus, anything that is not explicitly searched for will be missed. This greatly limits the flexibility and universal application of these techniques. We investigated the application of a rapid universal bacterial molecular identification method, IS-pro, to routine patient samples received in a clinical microbiology laboratory. IS-pro is a eubacterial technique based on the detection and categorization of 16S-23S rRNA gene interspace regions with lengths that are specific for each microbial species. As this is an open technique, clinicians do not need to decide in advance what to look for. We compared routine culture to IS-pro using 66 samples sent in for routine bacterial diagnostic testing. The samples were obtained from patients with infections in normally sterile sites (without a resident microbiota). The results were identical in 20 (30%) samples, IS-pro detected more bacterial species than culture in 31 (47%) samples, and five of the 10 culture-negative samples were positive with IS-pro. The case histories of the five patients from whom these culture-negative/IS-pro-positive samples were obtained suggest that the IS-pro findings are highly clinically relevant. Our findings indicate that an open molecular approach, such as IS-pro, may have a high added value for clinical practice.