Specific labeling of outer membrane vesicles with antibiotic-conjugated probe reveals early bacterial infections in blood

Qianbei Li, Zihao Ou (), Jinduan Lin, Die Tang, Bairong He, Yuanyuan Wu, Xinyue Huang, Xixin Huang, Bingbing Ru, Qianwen Wang, Weirong Yao, Bo Situ () and Lei Zheng ()
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Qianbei Li: Southern Medical University
Zihao Ou: Southern Medical University
Jinduan Lin: Guangzhou Medical University (Qingyuan People’s Hospital)
Die Tang: Medical Laboratory of the Third Affiliated Hospital of Shenzhen University
Bairong He: Southern Medical University
Yuanyuan Wu: Southern Medical University
Xinyue Huang: University of Electronic Science and Technology of China
Xixin Huang: Southern Medical University
Bingbing Ru: Southern Medical University
Qianwen Wang: Southern Medical University
Weirong Yao: The Second Hospital of Zhangzhou
Bo Situ: Southern Medical University
Lei Zheng: Southern Medical University

Nature Communications, 2025, vol. 16, issue 1, 1-14

Abstract: Abstract Bacterial outer membrane vesicles (OMVs) are nano-sized structures derived from the outer membrane of Gram-negative bacteria, which have emerged as key players in host-pathogen interactions, yet their potential as biomarkers remains largely unexplored due to the difficulty of identification in complex biological samples. Here we show an approach for detecting and quantifying bacterial OMVs in blood using a Polymyxin B-fluorescein probe (PmBF), which targets bacterial lipopolysaccharides (LPS). The probe selectively labels OMVs, enabling their differentiation from host extracellular vesicles and quantitative analysis using nano-flow cytometry. In male mouse models of pneumonia, we observe elevated serum PmBF+ EVs as early as 6 h post-infection, preceding positive blood cultures. In clinical samples, PmBF+ EVs show superior performance for diagnosing bacterial infections and differentiate them from virus or mycoplasma infections. Our findings highlight circulating PmBF+ EVs as promising biomarkers of bacterial infections.

Date: 2025
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DOI: 10.1038/s41467-025-58676-8

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