Molecular detection of culture-confirmed bacterial bloodstream infections with limited enrichment time.
Molecular detection of culture-confirmed bacterial bloodstream infections with limited enrichment time.
复制标题
在有限的富集时间内对培养证实的细菌血流感染进行分子检测。
DOI：
10.1128/jcm.01981-13
复制
发表时间：
2013
影响因子：
9.4
通讯作者：
Jordan,JeanneA
中科院分区：
文献类型：
--
作者：
Moore,MirandaS;McCann,ChaseD;Jordan,JeanneA
使用具有表型鉴定的自动化仪器的常规血液培养需要大量的时间来产生结果。本研究调查了使用PCR和焦磷酸测序生成结果的速度和准确性，并与获得革兰氏染色结果和培养确认血流感染病例的最终培养鉴定所需的时间进行了比较。同时进行研究和医生要求的血培养。在5 - 8 h、24 h和第5天，每小时取出孵育研究血培养液的等分试样。从这6个时间点的等分试样中提取DNA，并通过PCR和焦磷酸测序分析细菌rRNA基因靶标。然后将这些结果与医生要求的血培养结果进行比较。PCR和焦磷酸测序在5.8 ± 2.9 h的平均富集时间后准确识别了92%的所有培养确诊病例。当PCR和焦磷酸测序方案包括完成样本处理所需的时间时，分子方法在11.8 ± 2.9小时内产生结果，而获得革兰氏染色结果的平均时间为27.9 ± 13.6小时，生成最终基于培养的鉴定的平均时间为81.6 ± 24.0小时。分子方法能够准确检测孵育血培养瓶中存在的大多数细菌，平均比革兰氏染色结果快约16小时，比表型鉴定输入实验室信息系统快约3天。如果实施，这种更快速的分子方法可以最大限度地减少对患者使用不必要或无效抗生素的剂量。
Conventional blood culturing using automated instrumentation with phenotypic identification requires a significant amount of time to generate results. This study investigated the speed and accuracy of results generated using PCR and pyrosequencing compared to the time required to obtain Gram stain results and final culture identification for cases of culture-confirmed bloodstream infections. Research and physician-ordered blood cultures were drawn concurrently. Aliquots of the incubating research blood culture fluid were removed hourly between 5 and 8 h, at 24 h, and again at 5 days. DNA was extracted from these 6 time point aliquots and analyzed by PCR and pyrosequencing for bacterial rRNA gene targets. These results were then compared to those of the physician-ordered blood culture. PCR and pyrosequencing accurately identified 92% of all culture-confirmed cases after a mean enrichment time of 5.8 ± 2.9 h. When the time needed to complete sample processing was included for PCR and pyrosequencing protocols, the molecular approach yielded results in 11.8 ± 2.9 h compared to means of 27.9 ± 13.6 h to obtain the Gram stain results and 81.6 ± 24.0 h to generate the final culture-based identification. The molecular approach enabled accurate detection of most bacteria present in incubating blood culture bottles on average about 16 h sooner than Gram stain results became available and approximately 3 days sooner than the phenotypic identification was entered in the Laboratory Information System. If implemented, this more rapid molecular approach could minimize the number of doses of unnecessary or ineffective antibiotics administered to patients.