Quantitative PCR (qPCR) assays are the gold standard for diagnosis of Pneumocystis jirovecii pneumonia (PCP). However, they are laborious and require skilled personnel. Therefore, execution outside regular working hours of the molecular biology laboratory is limited. The eazyplex P. jirovecii assay (PJA) uses loop-mediated isothermal amplification for detection of P. jirovecii. It is performed directly with respiratory specimens, without the need for special skills, and delivers a result within 3 to 25 min. The goal of our study was to compare the performance of the eazyplex PJA with that of established P. jirovecii qPCR assays. All archived bronchoalveolar lavage fluid (BALF) samples that had previously tested positive for P. jirovecii by qPCR assay and 50 control samples (retrospective part), as well as all BALF samples received for P. jirovecii analysis over a period of 4 months (prospective part), were tested. Forty-nine patients with proven PCP and 126 patients without PCP were included. The sensitivity and specificity of the eazyplex PJA (95.7% and 96.5%, respectively) were comparable to those for three different P. jirovecii qPCR assays. The detection limit of the eazyplex PJA was analogous to 10³ copies of the major surface glycoprotein gene per 25 μl of BALF, corresponding to 10 to 20 P. jirovecii cells. The eazyplex PJA reliably discriminated patients with PCP from patients with P. jirovecii colonization. It delivered a positive result within a mean of 9 min 38 s and required a hands-on time of 2 min 45 s. In summary, the eazyplex PJA showed identical performance for the diagnosis of PCP, compared to qPCR assays. However, in terms of time to result, practicability, and robustness, the eazyplex PJA is clearly superior and allows for around-the-clock molecular testing.

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评价Amplexeazyplex®环介导的等温扩增测定法，以快速诊断吉氏肺孢子虫肺炎。

定量PCR（qPCR）分析是诊断吉氏肺孢子虫肺炎（PCP）的金标准。但是，它们费力并且需要熟练的人员。因此，限制了分子生物学实验室正常工作时间之外的执行。eazyplex P. jirovecii分析（PJA）使用回路介导的等温扩增来检测P. jirovecii。它直接对呼吸道标本进行，无需特殊技能，并在3至25分钟内产生结果。我们研究的目的是比较与既定的eazyplex PJA的性能P.卡氏肺囊虫定量PCR检测。所有先前测试为阳性的已归档的支气管肺泡灌洗液（BALF）样品通过qPCR分析对吉罗威氏假单胞菌和50个对照样品（回顾性部分）进行了测试，并对在4个月内接受的所有用于吉罗韦氏假单胞菌分析的BALF样品（预期部分）进行了测试。包括四十九例经证实的PCP患者和126例未经PCP的患者。eazyplex PJA的敏感性和特异性（分别为95.7％和96.5％）可与三种不同的jirovecii qPCR分析相比。eazyplex PJA的检出限类似于每25μlBALF中的10 ³个主要表面糖蛋白基因拷贝，相当于10至20个jirovecii细胞。eazyplex PJA可靠地区分了P. jirovecii患者和PCP患者殖民化。它在9分钟38 s的平均时间内提供了积极的结果，并且需要2分钟45 s的动手时间。总之，与qPCR分析相比，eazyplex PJA在诊断PCP方面表现出相同的性能。但是，就生成时间，实用性和鲁棒性而言，eazyplex PJA明显优越，并且可以进行全天候的分子测试。