Detection and Identification of Ehrlichia Species in Blood by Use of PCR and Electrospray Ionization Mass Spectrometry
Detection and Identification of Ehrlichia Species in Blood by Use of PCR and Electrospray Ionization Mass Spectrometry
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DOI：
10.1128/jcm.01669-09
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发表时间：
2010-02-01
影响因子：
9.4
通讯作者：
Tang, Yi-Wei
中科院分区：
文献类型：
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作者：
Eshoo, Mark W.;Crowder, Chris D.;Tang, Yi-Wei
快速检测和鉴定埃利希体种类可改善疑似埃利希体病患者的临床结果。我们描述了一种采用多位点 PCR 和电喷雾电离质谱 (PCR/ESI-MS) 直接从血液样本中检测和识别埃利希体属的检测方法。将结果与用作诊断测定的比色微量滴定 PCR 酶免疫测定 (PCR-EIA) 的结果进行比较。 2008年5月1日至8月1日，范德比尔特大学医院从临床疑似埃利希体病患者采集的213份全血样本中，PCR/ESI-MS检测结果显示40份埃利希体阳性，阳性率为18.8%。与PCR-EIA相比，PCR/ESI-MS的敏感性、特异性以及阳性和阴性预测值分别为95.0%、98.8%、95.0%和98.8%。通过 PCR/ESI-MS 和 PCR-EIA 检测埃利希体呈阳性的 38 份标本在物种水平上得到进一步表征，两种检测结果 100% 一致。此外，通过PCR/ESI-MS从4个标本中检测到立克次体，并通过血清学和PCR-EIA回顾性证实。在三个样本中，PCR/ESI-MS 检测鉴定出铜绿假单胞菌、脑膜炎奈瑟菌和金黄色葡萄球菌；这些经培养和/或临床诊断证实具有临床相关性。从标本处理到结果报告，PCR/ESI-MS检测可在6小时内完成，为埃立克体病的诊断提供了又一种实验室工具。此外，该系统可以直接从血液样本中快速检测和识别其他病原体。
Rapid detection and identification of Ehrlichia species improves clinical outcome for patients suspected of ehrlichiosis. We describe an assay that employs multilocus PCR and electrospray ionization mass spectrometry (PCR/ESI-MS) to detect and identify Ehrlichia species directly from blood specimens. The results were compared to those of a colorimetric microtiter PCR enzyme immunoassay (PCR-EIA) used as a diagnostic assay. Among 213 whole-blood samples collected from patients who were clinically suspected of ehrlichiosis from 1 May to 1 August 2008 at Vanderbilt University Hospital, 40 were positive for an Ehrlichia species by PCR/ESI-MS, giving a positive rate of 18.8%. In comparison to the PCR-EIA, PCR/ESI-MS possessed a sensitivity, a specificity, and positive and negative predictive values of 95.0%, 98.8%, 95.0%, and 98.8%, respectively. The 38 specimens that were positive for Ehrlichia by both PCR/ESI-MS and the PCR-EIA were further characterized to the species level, with 100% agreement between the two assays. In addition, Rickettsia rickettsii was detected by PCR/ESI-MS from four specimens that were confirmed retrospectively by serology and PCR-EIA. In three specimens, the PCR/ESI-MS assay identified Pseudomonas aeruginosa, Neisseria meningitidis, and Staphylococcus aureus; these were confirmed by culture and/or clinical diagnosis as being clinically relevant. From specimen processing to result reporting, the PCR/ESI-MS assay can be completed within 6 h, providing another laboratory tool for the diagnosis of ehrlichiosis. Moreover, this system may provide rapid detection and identification of additional pathogens directly from blood specimens.