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经支气管肺泡灌洗的 PCR 联合电喷雾电离-质谱检测用于血液系统疾病患者侵袭性霉菌感染的诊断。

PCR with electrospray ionization-mass spectrometry on bronchoalveolar lavage for detection of invasive mold infections in hematological patients.

机构信息

Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden.

Department of Clinical Microbiology, Karolinska University Hospital, Stockholm, Sweden.

出版信息

PLoS One. 2019 Feb 22;14(2):e0212812. doi: 10.1371/journal.pone.0212812. eCollection 2019.

Abstract

Invasive mold infections are life-threatening complications in patients with hematological malignancies. Conventional microbiological methods for diagnosing invasive pulmonary mold infections have low sensitivity, and molecular methods are being developed. Detection of molds using PCR with a narrow spectrum has been reported, but data with broad-spectrum PCR are lacking. In this study, the diagnostic performance and utility of a broad-spectrum PCR (broad-spectrum PCR with subsequent electrospray ionization-mass spectrometry, PCR/ESI-MS) for detection of molds in bronchoalveolar lavage (BAL) in 27 hematological patients with a new pulmonary infiltrate was analyzed. Using the revised EORTC/MSG criteria, PCR/ESI-MS was the only positive microbiological test in patients with proven invasive mold infection (n = 2) and correctly identified all cases of probable invasive pulmonary aspergillosis (n = 5). In patients with a possible invasive mold infection (n = 5), PCR/ESI-MS was positive in three patients. Mucorales was identified with PCR/ESI-MS in four patients that were all culture negative. The PCR/ESI-MS results had a clinical impact on antifungal therapy in 12 (44%) of the patients: modification of treatment in 6 (22%) patients and discontinuation in 6 (22%) patients. This study provides proof of concept that routine use of a broad-spectrum PCR for molds in bronchoalveolar lavage in immunocompromised patients is sensitive, fast, and has an impact on clinical decision-making.

摘要

侵袭性霉菌感染是血液恶性肿瘤患者生命受到威胁的并发症。传统的微生物学方法诊断侵袭性肺部霉菌感染的灵敏度较低,因此正在开发分子方法。已经报道了使用窄谱 PCR 检测霉菌的方法,但缺乏广谱 PCR 的数据。在这项研究中,分析了广谱 PCR(广谱 PCR 后进行电喷雾电离 - 质谱分析,PCR/ESI-MS)检测 27 例新发肺部浸润的血液系统恶性肿瘤患者支气管肺泡灌洗液(BAL)中霉菌的诊断性能和实用性。使用修订后的 EORTC/MSG 标准,PCR/ESI-MS 是仅有的 2 例确诊侵袭性霉菌感染患者的阳性微生物学检测方法,并正确识别了所有 5 例可能侵袭性肺曲霉病的病例。在可能患有侵袭性霉菌感染的 5 例患者中,PCR/ESI-MS 在 3 例患者中呈阳性。PCR/ESI-MS 在 4 例均培养阴性的患者中鉴定出毛霉科。PCR/ESI-MS 结果对 12 名(44%)患者的抗真菌治疗产生了临床影响:6 名(22%)患者改变治疗方案,6 名(22%)患者停止治疗。这项研究提供了概念验证,即常规使用广谱 PCR 检测免疫功能低下患者的支气管肺泡灌洗液中的霉菌具有敏感性、快速性,并对临床决策产生影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/976e/6386253/c583dd2de369/pone.0212812.g001.jpg

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