Multiplex Real-Time PCR for Respiratory Pathogens

Bacterial and viral DNAs as well as viral RNA were extracted from the nasopharyngeal swabs using RTP® Pathogen kit (STRATEC Molecular, Germany). A multiplex real-time PCR assay allowing the identification of 22 respiratory pathogens (Fast-track Diagnostics respiratory pathogens kit, FTD Luxembourg) was used as previously described [13 (link)]. Following nucleic extraction of EDTA-whole blood samples, another multiplex real-time PCR assay was performed to identify Staphylococcus aureus, Streptococcus pneumoniae and Haemophilus influenzae type b as previously described [14 (link)]. All RNA/DNA amplifications and detections were done using Bio-Rad CFX96 machine (Bio-Rad, USA).