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,...
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MDPI AG
2025-07-01
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| Online Access: | https://www.mdpi.com/2076-0817/14/7/692 |
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| author | Valeria Conciatori Sarah Di Sopra Elisa Franchin Ioannis Bekas Giuseppe Di Pietra Ignazio Castagliuolo Cristiano Salata Claudia Del Vecchio |
| author_facet | Valeria Conciatori Sarah Di Sopra Elisa Franchin Ioannis Bekas Giuseppe Di Pietra Ignazio Castagliuolo Cristiano Salata Claudia Del Vecchio |
| author_sort | Valeria Conciatori |
| collection | DOAJ |
| description | <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. |
| format | Article |
| id | doaj-art-0d5495be5f49481f937ba530d26bd507 |
| institution | DOAJ |
| issn | 2076-0817 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pathogens |
| spelling | doaj-art-0d5495be5f49481f937ba530d26bd5072025-08-20T03:08:13ZengMDPI AGPathogens2076-08172025-07-0114769210.3390/pathogens14070692Implementation of a Laboratory-Developed Test for the Diagnosis of <i>Mycoplasma pneumoniae</i> Using a High-Throughput ApproachValeria Conciatori0Sarah Di Sopra1Elisa Franchin2Ioannis Bekas3Giuseppe Di Pietra4Ignazio Castagliuolo5Cristiano Salata6Claudia Del Vecchio7Department of Molecular Medicine, University of Padua, 35121 Padua, ItalyDepartment of Molecular Medicine, University of Padua, 35121 Padua, ItalyDepartment of Molecular Medicine, University of Padua, 35121 Padua, ItalyDepartment of Molecular Medicine, University of Padua, 35121 Padua, ItalyUOC Medicina di Laboratorio, Ospedale San Bassano, ULSS 7 Pedemontana, Bassano del Grappa, 36061 Vicenza, ItalyDepartment of Molecular Medicine, University of Padua, 35121 Padua, ItalyDepartment of Molecular Medicine, University of Padua, 35121 Padua, ItalyDepartment of Molecular Medicine, University of Padua, 35121 Padua, Italy<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.https://www.mdpi.com/2076-0817/14/7/692<i>Mycoplasma pneumoniae</i>Panther Fusion<sup>®</sup> systemQuant Studio™5respiratory infectionsmolecular diagnostics |
| spellingShingle | Valeria Conciatori Sarah Di Sopra Elisa Franchin Ioannis Bekas Giuseppe Di Pietra Ignazio Castagliuolo Cristiano Salata Claudia Del Vecchio Implementation of a Laboratory-Developed Test for the Diagnosis of <i>Mycoplasma pneumoniae</i> Using a High-Throughput Approach Pathogens <i>Mycoplasma pneumoniae</i> Panther Fusion<sup>®</sup> system Quant Studio™5 respiratory infections molecular diagnostics |
| title | Implementation of a Laboratory-Developed Test for the Diagnosis of <i>Mycoplasma pneumoniae</i> Using a High-Throughput Approach |
| title_full | Implementation of a Laboratory-Developed Test for the Diagnosis of <i>Mycoplasma pneumoniae</i> Using a High-Throughput Approach |
| title_fullStr | Implementation of a Laboratory-Developed Test for the Diagnosis of <i>Mycoplasma pneumoniae</i> Using a High-Throughput Approach |
| title_full_unstemmed | Implementation of a Laboratory-Developed Test for the Diagnosis of <i>Mycoplasma pneumoniae</i> Using a High-Throughput Approach |
| title_short | Implementation of a Laboratory-Developed Test for the Diagnosis of <i>Mycoplasma pneumoniae</i> Using a High-Throughput Approach |
| title_sort | implementation of a laboratory developed test for the diagnosis of i mycoplasma pneumoniae i using a high throughput approach |
| topic | <i>Mycoplasma pneumoniae</i> Panther Fusion<sup>®</sup> system Quant Studio™5 respiratory infections molecular diagnostics |
| url | https://www.mdpi.com/2076-0817/14/7/692 |
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