Arcenas Rodney C, Uhl James R, Buckwalter Seanne P, Limper Andrew H, Crino Dana, Roberts Glenn D, Wengenack Nancy L
Division of Clinical Microbiology, Mayo Clinic College of Medicine, Rochester, MN 55905, USA.
Diagn Microbiol Infect Dis. 2006 Mar;54(3):169-75. doi: 10.1016/j.diagmicrobio.2005.08.006. Epub 2006 Jan 19.
Pneumocystis jiroveci is an important cause of pneumonia in immunocompromised individuals. This organism cannot be cultured, and therefore, diagnosis relies on microscopic identification of the organism using stains or antibodies. Although simple, these tests are insensitive and require expertise for accurate interpretation. We developed a real-time polymerase chain reaction (PCR) assay that provides sensitive and objective detection of Pneumocystis from bronchoalveolar lavage fluid. Primers and fluorescence resonance energy transfer probes were developed that target the cdc2 gene of P. jiroveci. Assay sensitivity is 6 copies of target per microliter of sample. No cross-reactivity occurs with other pathogens, and the PCR assay has a 21% increase in clinical sensitivity as compared with Calcofluor white staining. The real-time PCR assay provides a sensitive, rapid, and objective method for the detection of Pneumocystis from bronchoalveolar lavage fluid.
耶氏肺孢子菌是免疫功能低下个体肺炎的重要病因。该病原体无法培养，因此，诊断依赖于使用染色剂或抗体通过显微镜鉴定该病原体。尽管这些检测方法简单，但不敏感，且需要专业知识才能准确解读。我们开发了一种实时聚合酶链反应（PCR）检测方法，可对支气管肺泡灌洗 fluid 中的肺孢子菌进行灵敏且客观的检测。开发了针对耶氏肺孢子菌 cdc2 基因的引物和荧光共振能量转移探针。检测灵敏度为每微升样品 6 个靶标拷贝。与其他病原体无交叉反应，与荧光钙白染色相比，PCR 检测的临床灵敏度提高了 21%。实时 PCR 检测为从支气管肺泡灌洗 fluid 中检测肺孢子菌提供了一种灵敏、快速且客观的方法。
