Conciatori Valeria, Di Sopra Sarah, Franchin Elisa, Bekas Ioannis, Di Pietra Giuseppe, Castagliuolo Ignazio, Salata Cristiano, Del Vecchio Claudia
Department of Molecular Medicine, University of Padua, 35121 Padua, Italy.
Microbiology and Virology Diagnostic Unit, Padua University Hospital, 35121 Padua, Italy.
Pathogens. 2025 Jul 14;14(7):692. doi: 10.3390/pathogens14070692.
is a significant causative agent of atypical pneumonia in both children and adults. Timely and accurate diagnosis is crucial for appropriate patient management. Conventional methods for detecting , such as culture and serology, exhibit several limitations regarding sensitivity, specificity, and turnaround time. In contrast, real-time PCR is considered the most reliable, rapid, and sensitive technique for the diagnosis of infection. In this study, we adapted and validated an in-house real-time PCR assay for use on the fully automated Panther Fusion System. The validation process included two artificial samples, five external quality controls, and sixty-two patient samples. We evaluated the performance in terms of precision, sensitivity, linearity, and analytical sensitivity, comparing it to the original in-house assay. The Panther Fusion System demonstrated a broad dynamic range (16-1.6 × 10 copies/reaction), a robust correlation (94%) with the in-house assay, and comparable sensitivity (46 copies/mL vs. 25 copies/mL). The concordance between the in-house real-time PCR and the Panther Fusion System was 100% for both clinical samples and external quality controls. The adaptation of the test to the Panther Fusion System enabled the inclusion of among the pathogens monitored for respiratory infection surveillance. Throughout 2024, we analyzed 2567 samples, with a peak positivity rate of 38% observed in August. These findings underscore the significance of employing the diagnostic assay on the Panther Fusion System which proves valuable for the detection of infections. This platform offers the advantages of increased automation and greater throughput potential compared to other platforms, enhancing the efficiency of respiratory pathogen detection in clinical settings.
是儿童和成人非典型肺炎的重要病原体。及时准确的诊断对于恰当的患者管理至关重要。检测的传统方法，如培养和血清学，在敏感性、特异性和周转时间方面存在若干局限性。相比之下，实时聚合酶链反应（PCR）被认为是诊断感染最可靠、快速且敏感的技术。在本研究中，我们对一种内部实时PCR检测方法进行了调整并在全自动的Panther Fusion系统上进行了验证。验证过程包括两个人工样本、五个外部质量控制样本和62个患者样本。我们从精密度、灵敏度、线性和分析灵敏度方面评估了其性能，并与原始的内部检测方法进行了比较。Panther Fusion系统显示出较宽的动态范围（16 - 1.6×10拷贝/反应），与内部检测方法有很强的相关性（94%），且灵敏度相当（46拷贝/毫升对25拷贝/毫升）。内部实时PCR与Panther Fusion系统在临床样本和外部质量控制样本方面的一致性均为100%。将该检测方法适配到Panther Fusion系统使得能够将纳入呼吸道感染监测所监测的病原体之中。在2024年全年，我们分析了2567个样本，8月份观察到的阳性率峰值为38%。这些发现强调了在Panther Fusion系统上采用诊断检测方法的重要性，该方法被证明对检测感染很有价值。与其他平台相比，这个平台具有更高的自动化程度和更大的通量潜力，提高了临床环境中呼吸道病原体检测的效率。
