Related Experiment Video
Updated: Jul 12, 2026
03:53
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
Rapid multiplex nested PCR for detection of respiratory viruses
W Y Lam1, Apple C M Yeung, Julian W Tang
1Department of Microbiology, The Chinese University of Hong Kong, 1/F Clinical Science Building, Prince of Wales Hospital, Shatin, New Territories, Hong Kong.
Journal of Clinical Microbiology
|September 7, 2007
Summary
Rapid multiplex PCR assays offer a more sensitive and faster way to diagnose respiratory infections by detecting 21 pathogens. This improved diagnostic yield allows for quicker treatment decisions.
Area of Science:
- Molecular Biology
- Infectious Diseases
- Clinical Diagnostics
Background:
- Respiratory tract infections present with similar symptoms, necessitating accurate laboratory diagnosis.
- Conventional diagnostic methods often lack the sensitivity and speed required for timely intervention.
Purpose of the Study:
- To develop and evaluate multiplex nested PCR assays for the simultaneous detection of 21 respiratory pathogens.
- To assess the sensitivity, specificity, and efficiency of these rapid assays compared to traditional methods.
Main Methods:
- Development of five multiplex nested PCR assay groups utilizing fast PCR technology.
- Simultaneous detection of influenza A and B viruses, parainfluenza viruses, RSV, rhinoviruses, enteroviruses, coronaviruses, metapneumoviruses, M. pneumoniae, C. pneumoniae, L. pneumophila, and adenoviruses.
- Evaluation using 303 clinical specimens.
Main Results:
- Multiplex nested PCR assays demonstrated high specificity and 100- to 1,000-fold increased sensitivity over conventional methods.
- Achieved an overall positive detection rate of 48.5%, significantly higher than virus isolation (20.1%) and immunofluorescence assay (13.5%).
- Detected non-cultivatable viruses, including rhinoviruses and coronaviruses, contributing to a 15.6% gain in detection rate.
Conclusions:
- Rapid multiplex nested PCR assays significantly improve diagnostic yield for respiratory infections.
- Enhanced sensitivity and speed enable prompt diagnostic and interventive actions.
- These assays are valuable tools for identifying a broad range of respiratory pathogens.
