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Updated: May 21, 2026
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Development of Multiplex Real-Time RT-qPCR Assays for the Detection of SARS-CoV-2, Influenza A/B, and MERS-CoV
Published on: November 10, 2023
Performance of a novel microarray multiplex PCR for the detection of 23 respiratory pathogens (SYMP-ARI study)
S Bierbaum1, N Königsfeld, N Besazza
1Department of Virology, Institute of Medical Microbiology and Hygiene, University Medical Center Freiburg, Hermann-Herder Str. 11, 79104 Freiburg, Germany.
Summary
A new multiplex polymerase chain reaction (PCR) suspension microarray for respiratory infections showed promise but had lower sensitivity than traditional PCR, particularly in adults. Further studies are needed to confirm its utility.
Area of Science:
- Clinical Microbiology
- Molecular Diagnostics
- Infectious Diseases
Background:
- Accurate pathogen identification is crucial for managing acute febrile respiratory tract infections.
- Current diagnostic methods can be time-consuming, delaying optimal patient care.
- Novel multiplex assays offer the potential for rapid, simultaneous detection of multiple pathogens.
Purpose of the Study:
- To evaluate a novel multiplex polymerase chain reaction (PCR) suspension microarray for detecting 19 viral and 4 atypical bacterial targets in respiratory samples.
- To compare the performance of the multiplex PCR assay against a comprehensive set of sensitive monoplex real-time PCR assays.
- To assess the diagnostic accuracy, including sensitivity and specificity, of the multiplex assay.
Main Methods:
- A novel multiplex PCR suspension microarray assay was developed and validated.
- A gold standard was established using sensitive monoplex real-time PCR assays for each target.
- A total of 300 prospectively collected clinical samples, along with archived samples, were analyzed using both methods.
Main Results:
- The multiplex PCR detected at least one target in 46% of samples, compared to 55% with monoplex PCR.
- Positivity rates were significantly higher in pediatric patients (74%) than in adults (18%) using the multiplex assay.
- Overall concordance between multiplex and monoplex PCR was 77%, with lower sensitivity observed for the multiplex assay, mainly due to low target concentrations.
Conclusions:
- The novel multiplex PCR suspension microarray assay demonstrated its principle for respiratory pathogen detection.
- The assay exhibited lower overall sensitivity compared to monoplex PCR, suggesting potential limitations in certain patient populations or for low-concentration targets.
- Further research with larger sample sizes is recommended to fully ascertain the sensitivity and specificity for all targets, especially for atypical bacteria.
