Department of Infectious Diseases, Israel Institute for Biological Research, Ness-Ziona, Israel.
Department of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness-Ziona, Israel.
J Clin Microbiol. 2018 Mar 26;56(4). doi: 10.1128/JCM.01479-17. Print 2018 Apr.
Multiplexed detection technologies are becoming increasingly important given the possibility of bioterrorism attacks, for which the range of suspected pathogens can vary considerably. In this work, we describe the use of Luminex MagPlex magnetic microspheres for the construction of two multiplexed diagnostic suspension arrays, enabling antibody-based detection of bacterial pathogens and their related disease biomarkers directly from blood cultures. The first 4-plex diagnostic array enabled the detection of both anthrax and plague infections using soluble disease biomarkers, including protective antigen (PA) and anthrax capsular antigen for anthrax detection and the capsular F1 and LcrV antigens for plague detection. The limits of detection (LODs) ranged between 0.5 and 5 ng/ml for the different antigens. The second 2-plex diagnostic array facilitated the detection of (LOD of 1 × 10 CFU/ml) and (LOD of 1 × 10 CFU/ml) from blood cultures. Inoculated, propagated blood cultures were processed (15 to 20 min) via 2 possible methodologies (Vacutainer or a simple centrifugation step), allowing the direct detection of bacteria in each sample, and the entire assay could be performed in 90 min. While detection of bacteria and soluble markers from blood cultures using PCR Luminex suspension arrays has been widely described, to our knowledge, this study is the first to demonstrate the utility of the Luminex system for the immunodetection of both bacteria and soluble markers directly from blood cultures. Targeting both the bacterial pathogens as well as two different disease biomarkers for each infection, we demonstrated the benefit of the multiplexed developed assays for enhanced, reliable detection. The presented arrays could easily be expanded to include antibodies for the detection of other pathogens of interest in hospitals or labs, demonstrating the applicability of this technology for the accurate detection and confirmation of a wide range of potential select agents.
多重检测技术变得越来越重要，因为生物恐怖袭击的可能性，其可疑病原体的范围可能有很大的不同。在这项工作中，我们描述了使用 Luminex MagPlex 磁性微球构建两个多重诊断悬浮阵列，从而能够直接从血液培养物中检测细菌病原体及其相关疾病生物标志物。第一个 4 plex 诊断阵列能够使用可溶性疾病生物标志物检测炭疽病和鼠疫感染，包括炭疽病检测中的保护性抗原 (PA) 和炭疽荚膜抗原，以及鼠疫检测中的荚膜 F1 和 LcrV 抗原。不同抗原的检测限 (LOD) 在 0.5 到 5ng/ml 之间。第二个 2 plex 诊断阵列促进了对 （检测限为 1×10CFU/ml）和 （检测限为 1×10CFU/ml）从血液培养物中的检测。接种、增殖的血液培养物通过 2 种可能的方法（Vacutainer 或简单的离心步骤）进行处理（15 至 20 分钟），允许直接检测每个样本中的细菌，整个测定可以在 90 分钟内完成。虽然使用 PCR Luminex 悬浮阵列从血液培养物中检测细菌和可溶性标记物已经被广泛描述，但据我们所知，这项研究首次证明了 Luminex 系统用于直接从血液培养物中免疫检测细菌和可溶性标记物的实用性。针对每种感染的两种不同疾病生物标志物以及两种细菌病原体，我们证明了开发的多重检测方法在增强、可靠检测方面的优势。该阵列可以很容易地扩展到包括用于检测医院或实验室中其他感兴趣病原体的抗体，证明了该技术用于准确检测和确认广泛潜在选择剂的适用性。
