Dejing Liu: Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Second Military Medical University Shanghai 200433 People's Republic of China. ORCID
Bo Hu: Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Second Military Medical University Shanghai 200433 People's Republic of China.
Dingfa Peng: Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Second Military Medical University Shanghai 200433 People's Republic of China.
Shan Lu: Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Second Military Medical University Shanghai 200433 People's Republic of China.
Shunxiang Gao: Eye & ENT Hospital, State Key Laboratory of Medical Neurobiology, Institutes of Brain Science and Collaborative Innovation Center for Brain Science, Shanghai Medical College, Fudan University Shanghai 200032 China.
Zhengang Li: Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Second Military Medical University Shanghai 200433 People's Republic of China.
Lianghua Wang: Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Second Military Medical University Shanghai 200433 People's Republic of China.
Binghua Jiao: Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Second Military Medical University Shanghai 200433 People's Republic of China.
is a ubiquitous marine bacterium that may cause rapid and deadly infection, threatening lives of people living around natural bodies of water, especially in coastal regions. However, traditional culture-based methods are time-consuming and unable to detect Viable But Non-Culturable (VBNC) cells. In this work, we isolated a batch of detection aptamers specifically binding to in all culture status. With traditional whole bacteria-SELEX (Systematic Evolution of Ligands by EXponential enrichment), flow cytometer analysis and imaging, we identify 18 candidates and validated two of them (V8 and V13) as applicable aptamers. Their truncated sequences also showed comparable performance. The dissociation constant (KD) value of V8 is shown to be as low as 11.22 ± 1.32 nM. Optimal aptamers V8 and V13 are also validated to be effective to detect different strains under different binding environments using flow cytometry. As for detection parameters, the LOD of the V8 from cytometry is 29.96 CFU mL, and the linear range is 10-5 × 10 CFU mL. This is the first case demonstrating that aptamers can detect the existence of VBNC bacteria as well as live bacteria.
