A phage-based luminescent reporter platform for rapid typing of multiple capsular types of Klebsiella pneumoniae.
5 affiliations
- 1Medical Research Center, Shandong First Medical University & Shandong Academy of Medical Sciences, Shandong, China.
- 2State Key Laboratory of Microbial Diversity and Innovative Utilization, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
- 3College of Life Science, University of Chinese Academy of Sciences, Beijing, China.
- 4School of Life Sciences, Yunnan University, Yunnan, China.
- 5Department of Laboratory Medicine, Peking University Third Hospital, Beijing, China.
Abstract
Klebsiella pneumoniae is an important opportunistic pathogen, and its capsular polysaccharides are key virulence factors. Capsular typing is important for epidemiological surveillance, clinical diagnosis, and therapy development; however, methods for capsule-targeted detection remain limited. Here, we engineered a modular bioluminescent reporter phage platform leveraging bacteriophage receptor-binding proteins (RBPs) for specific K. pneumoniae capsular typing. Initially, the K2-specific bacteriophage ΦRCIP0109 was engineered with the nluc reporter gene to generate ΦRCIP0109::nluc for K2 detection. Using ΦRCIP0109::nluc as the chassis, we engineered reporter phages by swapping RBPs, thereby expanding the detection range to four additional clinically important K-types (K64, K47, K1, and K57). Reporter phage specificity and sensitivity were evaluated against clinical isolates and in simulated polymicrobial environments. Directed evolution was employed to introduce RBP mutations for enhanced phage adsorption. These five reporter phages achieved 100% specificity against clinical isolates, detecting concentrations as low as 10 colony-forming units/mL within 3.5-5.5 h and successfully differentiating host strains in polymicrobial synthetic urine. Directed evolution of an RBP yielded a 10- to 100-fold increase in luminescence compared to that of the wild-type phage. Together, these advances establish a scalable platform that can be expanded to additional capsular types and the detection of other pathogens, representing a platform with clear potential for integration into point-of-care diagnostics, guiding targeted antimicrobial therapy and precision phage therapy.
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BibTeX
@article{song2026phagebased,
title = {{A phage-based luminescent reporter platform for rapid typing of multiple capsular types of Klebsiella pneumoniae.}},
author = {Yiyao Song and Shisong Jing and Yi Li and Yinglu Guo and Jiangqing Huang and Xianbiao Bi and Dawei Wei and Chao Wang and Gang Zhang and Jiajia Zheng and Zhongrui Ma and Jie Feng},
journal = {HLife},
year = {2026},
month = jun,
doi = {10.1016/j.hlife.2026.06.001},
url = {https://europepmc.org/article/MED/42904010},
note = {CC BY-NC-ND},
}
RIS
TY - JOUR
TI - A phage-based luminescent reporter platform for rapid typing of multiple capsular types of Klebsiella pneumoniae.
AU - Yiyao Song
AU - Shisong Jing
AU - Yi Li
AU - Yinglu Guo
AU - Jiangqing Huang
AU - Xianbiao Bi
AU - Dawei Wei
AU - Chao Wang
AU - Gang Zhang
AU - Jiajia Zheng
AU - Zhongrui Ma
AU - Jie Feng
PY - 2026
DA - 2026/06/16
JO - HLife
DO - 10.1016/j.hlife.2026.06.001
UR - https://europepmc.org/article/MED/42904010
AB - Klebsiella pneumoniae is an important opportunistic pathogen, and its capsular polysaccharides are key virulence factors. Capsular typing is important for epidemiological surveillance, clinical diagnosis, and therapy development; however, methods for capsule-targeted detection remain limited. Here, we engineered a modular bioluminescent reporter phage platform leveraging bacteriophage receptor-binding proteins (RBPs) for specific K. pneumoniae capsular typing. Initially, the K2-specific bacteriophage ΦRCIP0109 was engineered with the nluc reporter gene to generate ΦRCIP0109::nluc for K2 detection. Using ΦRCIP0109::nluc as the chassis, we engineered reporter phages by swapping RBPs, thereby expanding the detection range to four additional clinically important K-types (K64, K47, K1, and K57). Reporter phage specificity and sensitivity were evaluated against clinical isolates and in simulated polymicrobial environments. Directed evolution was employed to introduce RBP mutations for enhanced phage adsorption. These five reporter phages achieved 100% specificity against clinical isolates, detecting concentrations as low as 10 colony-forming units/mL within 3.5-5.5 h and successfully differentiating host strains in polymicrobial synthetic urine. Directed evolution of an RBP yielded a 10- to 100-fold increase in luminescence compared to that of the wild-type phage. Together, these advances establish a scalable platform that can be expanded to additional capsular types and the detection of other pathogens, representing a platform with clear potential for integration into point-of-care diagnostics, guiding targeted antimicrobial therapy and precision phage therapy.
SN - 2949-9283
ER -