Simultaneous detection of respiratory syncytial virus types A and B and influenza virus types A and B in community-acquired pneumonia by reverse transcription-multiplex PCR

Abstract

Background: Respiratory syncytial virus (RSV) types A and B and influenza A and B cause about 80–90% of viral lower respiratory tract infections. It is impossible to distinguish the cause of viral respiratory infections by their clinical presentation. Multiplex RT-PCR has a significant advantage in that it permits the simultaneous amplification of several viruses in a single reaction facilitating cost-effective diagnosis and perhaps improved clinical management.
Objectives: In this study, our aim was to determine the prevalence of influenza A and B, and RSV types A and B among children with CAP, by the use of the newly developed rapid, accurate, and pathogen-specific technique of multiplex RT-PCR in order to accomplish the greatest positive effect on patient care and health care costs.
Study methodology: This study is a cross-sectional study involving 24 children admitted to the Children’s Hospital of the Ain Shams University due to severe lower respiratory tract infection (LRTI). Clinical and radiological assessment of all patients was performed followed by the molecular analysis of both respiratory and blood samples of all enrolled patients simultaneously by the use of the newly available reverse transcription-multiplex PCR technique (RT-m PCR).
Results: Viral pneumonia was detected in one-third of enrolled patients (8/24), with the predominance of respiratory syncytial virus A (4/8), followed by influenza A virus (3/8) and influenza B virus (1/8) while no cases of respiratory syncytial virus B was detected. The same results were identified in both blood and respiratory specimens.
Conclusion: Reverse transcription-multiplex PCR technique – multiplex has a significant advantage in that it permits the simultaneous amplification of several viruses in a single reaction making it well suited for use in epidemiological studies and to improve etiology-directed clinical management of viral pneumonia.

Keywords: Viral pneumonia; LRTI; Children; Reverse transcription-multiplex PCR; RSV; Influenza viruses