适体
指数富集配体系统进化
嗜肺军团菌
军团菌
生物传感器
寡核苷酸
化学
军团病
微生物学
生物
计算生物学
分子生物学
细菌
生物化学
DNA
遗传学
核糖核酸
基因
作者
Aysha Shaukat,Amani Chrouda,Saima Sadaf,Fatimah S. Alhamlan,Khaled Alkattan,Mohammed Zourob
标识
DOI:10.1016/j.biosx.2023.100411
摘要
Fluoriobacter Bozemanae (F. bozemanae) can cause Legionnaires' disease. The conventional methods for the detection of F. bozemanae are laborious, costly and has to be performed in centralized labs by skilled personnel. Therefore, there is an urgent need for rapid detection methods for F. bozemanae. The present work reports for the first electrochemical aptasensor for the rapid and specific detection of the most virulent type of Legionellosis, Fluoriobacter Bozemanae (F. bozemanae). Firstly, high-affinity aptamer was selected against the F. bozemanae using cell-based systematic evolution of ligands by exponential enrichment (Cell SELEX). The aptamers selections were based on 10 rounds of cell-SELEX with live F. bozemanae cells, and the counter-selection was performed against five other closely related Legionella sub-species. The selected oligonucleotide sequences were characterized using the dissociation constant (Kd). Apt-FB6 sequence gave a Kd value as low as 9.516 nM. This selected aptamer was integrated with electrochemical impedance spectroscopy (EIS). The impedimetric aptasensor for F. bozemanae exhibited a detection limit down to 10 CFU/mL. Finally, the aptasensor was tested against Legionella anisa, Legionella micdadei, Fluoriobacter dumoffii, and Legionella pneumophila serogroup 1 and 3. The biosensor didn't show significant cross-reactivity with the closely-related sub-species of Legionella. The selected aptasensor can be further exploited for further biosensing applications for clinical, environmental, and diagnostic purposes.
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