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Key performance evaluation of commercialized multiplex rRT-PCR kits for respiratory viruses: implications for
Wanyu Feng1,2,3, Yuqing Chen1,2,3, Yanxi Han1,2,3
1National Center for Clinical Laboratories, Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, Beijing Hospital/National Center of Gerontology, Beijing, China.
Microbiology Spectrum
|October 29, 2024
Summary
Multiplex real-time reverse transcription polymerase chain reaction (rRT-PCR) kits show variable performance for detecting respiratory viruses. While most kits offer good sensitivity for common viruses, some struggle with less prevalent ones, though they can detect co-infections.
Area of Science:
- Clinical Virology
- Molecular Diagnostics
- Infectious Disease Epidemiology
Background:
- Viral respiratory tract infections (RTIs) are common and require accurate diagnosis for effective management.
- Multiplex real-time reverse transcription polymerase chain reaction (rRT-PCR) assays are widely used for diagnosing viral RTIs.
- Evaluating the analytical performance of these multiplex assays is crucial for their reliable application.
Purpose of the Study:
- To assess the analytical sensitivity and competitive interference of six common multiplex rRT-PCR kits for respiratory virus detection in China.
- To compare the performance of multiplex assays against singleplex kits for clinically significant respiratory viruses.
- To understand the diagnostic capabilities of multiplex rRT-PCR in identifying viral co-infections.
Main Methods:
- Evaluation of analytical sensitivity (limit of detection) for various respiratory viruses using six multiplex rRT-PCR kits.
- Assessment of competitive interference between different viral targets within multiplex assays.
- Comparison of multiplex assay performance with established singleplex methods.
Main Results:
- Significant variability in the limits of detection was observed across different viruses and kits.
- Most multiplex kits exhibited comparable or superior analytical sensitivity for major respiratory viruses like influenza and RSV.
- Multiplex kits showed reduced sensitivity for certain viruses such as human rhinovirus and parainfluenza virus.
- Most kits successfully detected co-infections, even with one analyte at a low concentration.
Conclusions:
- Multiplex rRT-PCR kits offer a valuable tool for diagnosing viral RTIs, with generally good performance for common pathogens.
- Optimization is needed for multiplex assays to improve sensitivity for less common respiratory viruses.
- These assays demonstrate utility in identifying complex co-infections, aiding clinical decision-making.
