Research and Development, ZEUS Scientific, Raritan, New Jersey, United States of America.
PLoS One. 2013 Oct 14;8(10):e78488. doi: 10.1371/journal.pone.0078488. eCollection 2013.
Surveillance of bloodstream infections (BSI) is a high priority within the hospital setting. Broth-based blood cultures are the current gold standard for detecting BSI, however they can require lengthy incubation periods prior to detection of positive samples. We set out to demonstrate the feasibility of using enzymatic template generation and amplification (ETGA)-mediated measurement of DNA polymerase activity to detect microbes from clinical blood cultures. In addition to routine-collected hospital blood cultures, one parallel aerobic blood culture was collected and immediately refrigerated until being transported for ETGA analysis. After refrigeration holding and transport, parallel-collected cultures were placed into a BACTEC incubator and ETGA time-course analysis was performed. Of the 308 clinical blood cultures received, 22 were BACTEC positive, and thus were initially selected for ETGA time course analysis. The ETGA assay detected microbial growth in all 22 parallel-positive blood cultures in less time than a BACTEC incubator and also yielded genomic DNA for qPCR-based organism identification. In summary, feasibility of detecting microbes from clinical blood culture samples using the ETGA blood culture assay was demonstrated. Additional studies are being considered towards development of clinically beneficial versions of this methodology.
血流感染（BSI）的监测在医院环境中是重中之重。基于肉汤的血培养是目前检测 BSI 的金标准，但在检测阳性样本之前，它们可能需要较长的孵育时间。我们旨在证明使用酶促模板生成和扩增（ETGA）介导的 DNA 聚合酶活性测量来检测临床血培养物中的微生物的可行性。除了常规收集的医院血培养物外，还收集了一个平行的需氧血培养物，并立即冷藏，直到进行 ETGA 分析。冷藏保存和运输后，将平行收集的培养物放入 BACTEC 孵育箱中，并进行 ETGA 时间过程分析。在收到的 308 份临床血培养物中，有 22 份 BACTEC 阳性，因此最初选择用于 ETGA 时间过程分析。ETGA 检测在少于 BACTEC 孵育箱的时间内检测到所有 22 个平行阳性血培养物中的微生物生长，并且还产生了基于 qPCR 的用于鉴定生物体的基因组 DNA。总之，使用 ETGA 血培养检测来自临床血培养物样本中的微生物的可行性得到了证明。正在考虑进一步研究以开发这种方法的临床有益版本。
