Nucleic acid detection and quantitative analysis of influenza virus using exhaled breath condensate

Exhaled breath condensate (EBC) is increasingly being used as a non-invasive method for disease diagnosis and environmental exposure assessment. We previously detected the nucleic acids of bacterial pathogens in EBC. Influenza viruses can be transmitted through aerosols during coughing and exhaling. Existing detection methods for influenza have various limitations. The EBC collection method is convenient, non-invasive, and reduces the risk of exposure. We investigated the detection of influenza virus in EBC using a sensitive nucleic acid testing method and performed quantitative analysis to evaluate the present and content of influenza virus in the breath. We evaluated 30 patients with respiratory tract infection during the 2019 influenza season. The clinical data and samples of nasal swabs were collected for rapid influenza diagnostic (antigen) tests. Pharyngeal swab and EBC samples were used for influenza virus nucleic acid detection. Each EBC sample was assessed twice as well as at one-month follow-up of the patients. The nucleic acid test in the EBC of 30 cases revealed 20 and two cases of influenza A and B, respectively, giving a detection rate of 73.3%. The rapid influenza diagnostic (antigen) tests revealed four and 12 cases of influenza A and B, respectively, with a detection rate of 53.3%. All pharyngeal swab samples evaluated by the nucleic acid test were influenza-positive; 12 cases were positive for both influenza A and B and 18 cases were positive for influenza B alone. The influenza viral load in the EBC was 10³-10⁷ copies ml^-1. Among the 16 patients followed-up after 1 month, 4 were positive (25%) in EBC samples and 10 were positive (62.5%) in pharyngeal swab samples. It was preliminary exploration that influenza virus could be detected in EBC. EBC is one of the sample types that would be used for molecular diagnosis of influenza.