Evaluation Of A Real-time PCR Assay For Diagnosis Of PCP Using Oral Washes

耶氏肺孢子虫是免疫功能低下个体肺炎（PCP）的重要原因。诊断PCP的标准方法是对诱导痰或支气管肺泡灌洗（BAL）样本制备的涂片进行显微镜检查。我们还开发了一种快速，定量PCR（聚合酶链反应）检测呼吸道样本中的P. jiroveci。由于PCR检测比显微镜检查更敏感，我们已经证明可以在口腔清洗液等非侵入性样本中检测到耶氏疟原虫。此外，该测试可以检测仅定植或具有亚临床感染的患者。 与加州大学旧金山分校弗朗西斯科（UCSF）的一项合作研究正在进行中，以尝试检测接触PCP患者后医护人员上呼吸道的耶氏疟原虫定植。已从60名医护人员中采集了口腔清洗液样本，并将通过PCR处理Jiroveci疟原虫。本研究的结果将确定这种重要病原体无症状定植的发生率。 第二项研究已经开始与合作者在旧金山弗朗西斯科总医院和穆拉戈医院工作人员在乌干达。口腔冲洗液收集自在医院结核病门诊就诊的呼吸道疾病患者。这些标本已被处理用于分枝杆菌感染（即，标本的抗酸涂片和培养） 与弗朗西斯科总医院，NIH临床中心CCM同事和乌干达Mulago医院工作人员合作开始了一项新的研究，从呼吸系统疾病患者中收集标本，以确定是否可以在口腔清洗液中检测到肺孢子虫。 标本已送至NIH，用于使用MSG肺孢子虫PCR系统进行研究。 迄今已收到100多个样本。

Pneumocystis jiroveci is an important cause of pneumonia (PCP) in immunocompromised individuals. The standard approach for diagnosing PCP is a microscopic examination of smears prepared from induced sputum or bronchial alveolar lavage (BAL) samples. We have also developed a rapid, quantitative PCR (polymerase chain reaction) assay for the detection of P. jiroveci in respiratory samples. Because the PCR assay is more sensitive that microscopic examination, we have demonstrated the P. jiroveci can be detected in noninvasive specimens such as oral washes. Additionally, the test can detect patient who are only colonized or have sub-clinical infections. A collaborative study with the University of California at San Francisco (UCSF) is underway to attempt to detect P. jiroveci colonization of the upper airways of healthcare workers after exposure to patients with PCP. Oral wash specimens have been collected from 60 healthcare workers and will be processed by PCR for P. jiroveci. The results of this study will define the incidence of asymptomatic colonization with this important pathogen. A second study has been initiated with collaborators at San Francisco General Hospital and Mulago Hospital workers in Uganda. Oral washes have been collected from patients with respiratory diseases who present with respiratory diseases at the hospital Tuberculosis Clinic. These specimens have been processed for mycobacterial infections (i.e., acid-fast smear and culture of the specimen) A new study was begun in collaboration with the San Francisco General Hospital, NIH Clinical Center CCM coworkers, and Mulago Hospital workers in Uganda to collect specimens from patients with respiratory diseases to determine if Pneumocystis jiroveci can be detected in oral washes. Specimens have been sent to the NIH for studies using the MSG Pneumocystis PCR system. Over 100 samples have been received to date.