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J Mol Diagn . Assessment of Multiplex Molecular Tests for Detecting Viral Infections in Lower Respiratory Tract Specimens

tetano

Editor, Senior Moderator
J Mol Diagn


. 2026 Apr 22:S1525-1578(26)00064-4.
doi: 10.1016/j.jmoldx.2026.03.008. Online ahead of print.
Assessment of Multiplex Molecular Tests for Detecting Viral Infections in Lower Respiratory Tract Specimens

Giuseppe Sberna[SUP] 1 [/SUP], Pierpaolo Paba[SUP] 2 [/SUP], Cosmina Mija[SUP] 3 [/SUP], Gaetana Costanza[SUP] 2 [/SUP], Fabiano Brillo[SUP] 3 [/SUP], Sandro Grelli[SUP] 4 [/SUP], Fabrizio Maggi[SUP] 3 [/SUP], Eleonora Lalle[SUP] 3 [/SUP]


Affiliations
Abstract

Lower respiratory tract infections (LRTIs) represent a major cause of morbidity and mortality, particularly among critically ill patients. Rapid molecular diagnostic tests have significantly improved the detection of respiratory pathogens. However, most commercial assays are validated exclusively for upper respiratory tract (URT) specimens, limiting their applicability for lower respiratory tract (LRT) samples, which are often more representative of disease in advanced or severe cases. This study evaluated the diagnostic performance of two commercially available assays, the "BIOFIRE Respiratory Panel 2.1 plus" and the "Panther Fusion SARS-CoV-2/Flu A/B/RSV assay", on bronchoalveolar lavage (BAL) specimens, using the "Allplex Respiratory Panel 1/2/3" and "Allplex SARS-CoV-2 Assay" as reference methods validated both for URT and LRT matrices. 132 BAL samples Were analyzed. Biofire identified more positives than Allplex, particularly for HRV/EV, HPIV, and non-SARS-CoV-2 coronaviruses. The overall agreement between Biofire and Allplex was fair (κ=0.237), and pathogen-specific concordance showed an almost perfect agreement for SARS-CoV-2 (κ=0.841), influenza A/B (κ=0.808), and HPIV (κ=0.884). The Panther assay showed substantial agreement with Allplex (κ=0.719) and near-perfect concordance for SARS-CoV-2 and influenza viruses, while Biofire and Panther exhibited almost perfect inter-assay agreement (κ=0.903). These findings demonstrate that assays validated for URT specimens can perform reliably on BAL samples, underscoring the diagnostic potential of LRT matrices and the need for expanded validation of molecular respiratory panels across specimen types.


 
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