Optimal sensitivity to detect low Pneumocystis loads is of importance to take individual and collective measures to avoid evolution towards Pneumocystis pneumonia and outbreaks in immunocompromised patients. This study compares two qPCR procedures, a new automated RTqPCR using the GeneLEAD VIII extractor/thermocycler (GLVIII; ∼2.2 h workflow) and a previously validated in-house qPCR assays (IH; ∼5 h workflow) both targeting mtSSU and mtLSU for detecting P. jirovecii in 213 respiratory samples. GLVIII was found to be more sensitive than IH, detecting eight more specimens. Bland-Altman analysis between the two procedures showed a Cq bias of 1.17 ± 0.07 in favor of GLVIII. Lay Summary The fungus Pneumocystis needs to be detected early in respiratory samples to prevent pneumonia in immunocompromised hosts. We evaluated a new commercial RTqPCR on 213 respiratory samples to detect Pneumocystis and found it more sensitive and faster than our routine sensitive in-house qPCR assay.

中文翻译：

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一种新的商业逆转录酶定量 PCR 提高了检测呼吸道标本中耶氏肺孢子菌的灵敏度

检测低肺孢子菌负荷的最佳灵敏度对于采取个体和集体措施以避免演变为肺孢子菌肺炎和免疫功能低下患者的爆发非常重要。本研究比较了两种 qPCR 程序，一种使用 GeneLEAD VIII 提取器/热循环仪（GLVIII；~2.2 小时工作流程）的新自动化 RTqPCR 和先前验证的内部 qPCR 测定（IH；~5 小时工作流程），均针对 mtSSU 和 mtLSU 进行检测213 个呼吸道样本中的 P. jirovecii。发现 GLVIII 比 IH 更敏感，可检测到 8 个以上的样本。两个程序之间的 Bland-Altman 分析显示 Cq 偏差为 1.17 ± 0.07，有利于 GLVIII。总结 需要尽早在呼吸道样本中检测到真菌肺孢子菌，以预防免疫功能低下的宿主发生肺炎。