Respiratory diseases are challenging to diagnose rapidly and accurately due to diverse pathogens, overlapping symptoms, and high infection rates. Here, we developed a suspension array based on magnetic microspheres encoded with blue fluorescent protein (mTagBFP) and Cy5-labeled anti-Tag ssDNA (MB@mTagBFP&anti-Tag-Cy5) for simultaneous detection of five core respiratory pathogens. Multiplex PCR was performed using 5'-Tag-labeled upstream primers and 5'-biotinylated downstream primers, yielding dual-labeled products (Tag and biotin). The resulting complexes (MB@mTagBFP&anti-Tag-Cy5-Tag-dsDNA-phycoerythrin) were analyzed by flow cytometry, achieving a detection limit of 1 copy/μL (3 SD/K). Linear quantification ranged from 101 to 105 copies/μL for respiratory syncytial virus and SARS-CoV-2, and from 101 to 106 copies/μL for influenza A, influenza B, and Mycoplasma pneumoniae. Recovery rates were 89.73-107.25%. Clinical testing of 100 samples yielded 97% sensitivity and 90% specificity, effectively distinguishing single and mixed infections. This integrated method enables rapid, high-throughput, and sensitive detection of respiratory pathogens, offering critical support for early diagnosis, treatment guidance, and epidemiological surveillance, with broad applications in infection control and disease prevention.
Dual color microsphere encoded by fluorescent protein and Cy5-labeled anti-Tag ssDNA for multiplex detection of respiratory pathogens.
发布日期:2026/10/27 14:04:47

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