A sensitive mass spectrometry-based method to identify common respiratory pathogens in children

AUTHOR AFFILIATIONSSee affiliation list on p. 8. 

ABSTRACT Public health threats posed by emerging respiratory infections are a  significant concern, particularly in children and infants. Traditional culture-based  detection methods are time-consuming and typically require 1–3 days. Herein, we  developed and evaluated a 23-plex common respiratory pathogen mass spectrometry  assay that enables the simultaneous detection of 18 common respiratory pathogens in  children. This assay combines matrix-assisted laser desorption/ionization time of flight mass spectrometry with multiplex reverse transcription-PCR and targets 11 bacterial and  7 viral pathogens (including 10 subtypes), and two internal controls. The detection limit  of the common respiratory pathogen mass spectrometry assay was as low as 1 copy/µL,  with no cross-reactivity with other organisms. We assessed the clinical performance of  the common respiratory pathogen mass spectrometry assay using respiratory samples  from 450 children. The total 450 clinical specimens underwent analysis via matrix-assisted laser desorption/ionization time of flight mass spectrometry, and the outcomes  were juxtaposed with those derived from real-time reverse-transcriptase PCR conducted  concurrently. The concordance between these methods was 96.0%, and the multiple  infection identification rate was 7.1%. This innovative approach enables the simultaneous analysis of numerous outcomes from a solitary examination across 192 specimens  within a timeframe of approximately 7 hours, with a dramatically reduced sample use  and cost. In summary, the common respiratory pathogen mass spectrometry assay  is a sensitive, accurate, and cost-effective method for detecting common respiratory  pathogens in children and has the potential to revolutionize the diagnosis of respiratory  tract infections. IMPORTANCE This study aimed to present and evaluate a novel co-detection method  that enables the simultaneous identification of 11 bacterial and 7 viral pathogens in  about 7 hours using matrix-assisted laser desorption/ionization time of flight mass  spectrometry. Our approach utilizes a combination of multiplex reverse transcriptionPCR and matrix-assisted laser desorption/ionization time of flight mass spectrometry,  which overcomes the limitations of conventional assays, which include a long assessment time, technical difficulty, and high costs. As a screening method for common  respiratory pathogens in children, common respiratory pathogen mass spectrometry  assay has the potential to revolutionize the diagnosis of respiratory tract infections by  providing an accurate etiological diagnosis. The common respiratory pathogen mass  spectrometry assay is expected to be a critical tool for the diagnosis of respiratory  infections in children, offering a more efficient, cost-effective, and accurate approach for  the detection of common respiratory pathogens.

KEYWORDS children common respiratory pathogen, matrix-assisted laser desorption/ionization time of flight mass spectrometry, pediatric respiratory tract infections,  multiplex reverse transcription-PCR, multipathogen infections