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Published on: November 28, 2016
Implementation of a Laboratory-Developed Test for the Diagnosis of Mycoplasma pneumoniae Using a High-Throughput
Valeria Conciatori1, Sarah Di Sopra1, Elisa Franchin1,2
1Department of Molecular Medicine, University of Padua, 35121 Padua, Italy.
Abstract:
Mycoplasma pneumoniae 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 M. pneumoniae, 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 M. pneumoniae 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 × 107 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 M. pneumoniae 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 M. pneumoniae diagnostic assay on the Panther Fusion® System which proves valuable for the detection of M. pneumoniae 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.
Insights
Accurate diagnosis of Mycoplasma pneumoniae pneumonia is vital. An automated Panther Fusion system provides reliable, sensitive, and rapid detection, improving respiratory infection surveillance and patient care.
Area of Science:
- Clinical microbiology
- Molecular diagnostics
- Infectious disease epidemiology
Background:
- Mycoplasma pneumoniae causes atypical pneumonia in all age groups.
- Conventional diagnostic methods have limitations in speed, sensitivity, and specificity.
- Real-time PCR offers a more reliable and rapid diagnostic approach.
Purpose of the Study:
- To adapt and validate an in-house real-time PCR assay for Mycoplasma pneumoniae on the automated Panther Fusion system.
- To evaluate the performance characteristics of the adapted assay.
- To assess the utility of the Panther Fusion system for respiratory pathogen surveillance.
Main Methods:
- Validation using artificial samples, external quality controls, and patient samples.
- Performance evaluation included precision, sensitivity, linearity, and analytical sensitivity.
- Comparison with the original in-house real-time PCR assay.
Main Results:
- The Panther Fusion system showed a broad dynamic range (16-1.6 × 10^7 copies/reaction).
- A 94% correlation and 100% concordance were observed with the in-house assay.
- Comparable sensitivity was achieved (46 copies/mL vs. 25 copies/mL).
Conclusions:
- The automated Panther Fusion system is a valuable platform for accurate Mycoplasma pneumoniae detection.
- This adaptation enhances automation and throughput for respiratory pathogen surveillance.
- The system improves the efficiency of diagnosing Mycoplasma pneumoniae infections in clinical settings.
