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Author Spotlight: Advancements in Multiplex Detection of Respiratory Viruses
Published on: November 10, 2023
Detection of respiratory viruses and bacteria in children using a twenty-two target reverse-transcription real-time
Chelsey Ellis1, Amita Misir1, Charles Hui1
1Children's Hospital of Eastern Ontario, Children's Hospital of Eastern Ontario Research Institute, University of Ottawa, Ottawa, Canada.
Insights
A new ready-to-use respiratory infection reverse-transcription real-time polymerase chain reaction (RT-qPCR) panel significantly improved pathogen detection in children. This rapid test identified more viruses and bacteria compared to standard methods.
Area of Science:
- Clinical microbiology
- Molecular diagnostics
- Pediatric infectious diseases
Background:
- Rapid detection of pediatric respiratory pathogens is crucial for effective patient care and antibiotic stewardship.
- Current diagnostic methods may not capture the full spectrum of causative agents.
- A comprehensive, ready-to-use panel could enhance diagnostic capabilities.
Purpose of the Study:
- To design and evaluate a 22-target respiratory infection reverse-transcription real-time polymerase chain reaction (RT-qPCR) panel.
- To assess the panel's performance in detecting respiratory pathogens in pediatric nasopharyngeal aspirate (NPA) samples.
- To compare the panel's detection rate with existing hospital laboratory methods.
Main Methods:
- A 22-target RT-qPCR panel with dried-down assays was developed for 18 respiratory viruses and 4 bacteria.
- Nasopharyngeal aspirate (NPA) samples underwent automated nucleic acid extraction followed by rapid qPCR.
- RT-qPCR results were compared against the hospital's standard diagnostic testing methods.
Main Results:
- The RT-qPCR panel detected one or more pathogens in 83% of 159 pediatric NPA samples, significantly higher than standard methods (59%).
- Improved detection was noted for rhinoviruses, parainfluenza viruses, bocavirus, and coronaviruses.
- The panel's internal control remained stable with room temperature storage for two months.
Conclusions:
- The RT-qPCR panel effectively identified a high proportion of respiratory pathogens in pediatric samples.
- The pre-made, ready-to-use panel offered rapid results (approx. 90 minutes) and ease of use.
- Further studies are planned to evaluate the panel's impact on patient care and antibiotic stewardship.
Background:
Rapid detection of the wide range of viruses and bacteria that cause respiratory infection in children is important for patient care and antibiotic stewardship. We therefore designed and evaluated a ready-to-use 22 target respiratory infection reverse-transcription real-time polymerase chain reaction (RT-qPCR) panel to determine if this would improve detection of these agents at our pediatric hospital.
Methods:
RT-qPCR assays for twenty-two target organisms were dried-down in individual wells of 96 well plates and saved at room temperature. Targets included 18 respiratory viruses and 4 bacteria. After automated nucleic acid extraction of nasopharyngeal aspirate (NPA) samples, rapid qPCR was performed. RT-qPCR results were compared with those obtained by the testing methods used at our hospital laboratories.
Results:
One hundred fifty-nine pediatric NPA samples were tested with the RT-qPCR panel. One or more respiratory pathogens were detected in 132/159 (83%) samples. This was significantly higher than the detection rate of standard methods (94/159, 59%) (P<0.001). This difference was mainly due to improved RT-qPCR detection of rhinoviruses, parainfluenza viruses, bocavirus, and coronaviruses. The panel internal control assay performance remained stable at room temperature storage over a two-month testing period.
Conclusion:
The RT-qPCR panel was able to identify pathogens in a high proportion of respiratory samples. The panel detected more positive specimens than the methods in use at our hospital. The pre-made panel format was easy to use and rapid, with results available in approximately 90 minutes. We now plan to determine if use of this panel improves patient care and antibiotic stewardship.
