Implementation of a Laboratory-Developed Test for the Diagnosis of <i>Mycoplasma pneumoniae</i> Using a High-Throughput Approach

<i>Mycoplasma pneumoniae</i> 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 <i>M. pneumoniae</i>, such as culture and serology,...

Full description

Saved in:
Bibliographic Details
Main Authors: Valeria Conciatori, Sarah Di Sopra, Elisa Franchin, Ioannis Bekas, Giuseppe Di Pietra, Ignazio Castagliuolo, Cristiano Salata, Claudia Del Vecchio
Format: Article
Language:English
Published: MDPI AG 2025-07-01
Series:Pathogens
Subjects:
Online Access:https://www.mdpi.com/2076-0817/14/7/692
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:<i>Mycoplasma pneumoniae</i> 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 <i>M. pneumoniae</i>, 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 <i>M. pneumoniae</i> infection. In this study, we adapted and validated an in-house real-time PCR assay for use on the fully automated Panther Fusion<sup>®</sup> 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<sup>®</sup> System demonstrated a broad dynamic range (16–1.6 × 10<sup>7</sup> 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<sup>®</sup> System was 100% for both clinical samples and external quality controls. The adaptation of the test to the Panther Fusion<sup>®</sup> System enabled the inclusion of <i>M. pneumoniae</i> 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 <i>M. pneumoniae</i> diagnostic assay on the Panther Fusion<sup>®</sup> System which proves valuable for the detection of <i>M. pneumoniae</i> 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.
ISSN:2076-0817