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评价两种商业实时 PCR 试剂盒在侵袭性肺曲霉病患者支气管肺泡灌洗液中曲霉 DNA 检测的应用。

Evaluation of Two Commercial Real-Time PCR Kits for Aspergillus DNA Detection in Bronchoalveolar Lavage Fluid in Patients with Invasive Pulmonary Aspergillosis.

机构信息

Laboratoire de Parasitologie et de Mycologie Médicale, Hôpitaux Universitaires de Strasbourg, Strasbourg, France.

Laboratoire de Parasitologie et de Mycologie Médicale, Hôpitaux Universitaires de Strasbourg, Strasbourg, France; Institut de Parasitologie et de Pathologie Tropicale, Fédération de Médecine Translationnelle, Université de Strasbourg, Strasbourg, France.

出版信息

J Mol Diagn. 2018 May;20(3):298-306. doi: 10.1016/j.jmoldx.2017.12.005. Epub 2018 Feb 19.

Abstract

Invasive pulmonary aspergillosis (IPA) is a common complication of immunosuppression. Rapid diagnosis using molecular techniques is essential to improve patient survival. PCR techniques are promising in enhancing Aspergillus detection in blood and respiratory samples. We evaluate for the first time the performances of two commercial real-time PCR kits, the A. fumigatus Bio-Evolution and the MycoGENIE A. fumigatus for the detection of A. fumigatus DNA in bronchoalveolar lavage (BAL) from patients with and without IPA. Seventy-three BAL samples were included. Thirty-one of them corresponded to patients with probable IPA, 11 to patients with possible IPA, and 31 to patients without aspergillosis, according to the 2008 European Organization for Research and Treatment of Cancer/Mycoses Study Group criteria. In the probable IPA group, A. fumigatus Bio-Evolution and the MycoGENIE A. fumigatus real-time PCR kits showed a specificity of 100% and a sensitivity of 81% and 71%, respectively. The A. fumigatus Bio-Evolution detected Aspergillus DNA in the 14 BAL samples with a positive Aspergillus culture result, whereas the MycoGENIE A. fumigatus PCR result was positive only for 12. In the possible IPA group, there were no positive real-time PCR or positive Aspergillus culture results. For the patients without aspergillosis, no positive result was observed for real-time PCR kit, despite the presence of various other non-Aspergillus pathogens in this group. Our study demonstrates an excellent specificity and a good sensitivity of A. fumigatus DNA detection in BAL samples with both kits.

摘要

侵袭性肺曲霉病(IPA)是免疫抑制的常见并发症。使用分子技术进行快速诊断对于提高患者生存率至关重要。PCR 技术在提高血液和呼吸道样本中曲霉属检测方面具有广阔的前景。我们首次评估了两种商业实时 PCR 试剂盒,即 A. fumigatus Bio-Evolution 和 MycoGENIE A. fumigatus,用于检测有无 IPA 的患者支气管肺泡灌洗液(BAL)中的 A. fumigatus DNA。共纳入 73 份 BAL 样本。根据 2008 年欧洲癌症研究与治疗组织/真菌病研究组标准,其中 31 份来自疑似 IPA 患者,11 份来自可能 IPA 患者,31 份来自无曲霉病患者。在疑似 IPA 组中,A. fumigatus Bio-Evolution 和 MycoGENIE A. fumigatus 实时 PCR 试剂盒的特异性均为 100%,敏感性分别为 81%和 71%。A. fumigatus Bio-Evolution 检测到 14 份阳性曲霉培养结果的 BAL 样本中的曲霉 DNA,而 MycoGENIE A. fumigatus PCR 结果仅为 12 份阳性。在可能 IPA 组中,没有实时 PCR 或阳性曲霉培养结果。对于无曲霉病患者,实时 PCR 试剂盒均未检测到阳性结果,尽管该组存在各种其他非曲霉病原体。我们的研究表明,这两种试剂盒均能在 BAL 样本中出色地特异性检测到 A. fumigatus DNA,且具有良好的敏感性。

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