Aptamer-Based Impedimetric Sensor for Bacterial Typing

被引:65
作者
Labib, Mahmoud [1 ]
Zamay, Anna S. [2 ]
Kolovskaya, Olga S. [2 ]
Reshetneva, Irina T. [3 ]
Zamay, Galina S. [2 ]
Kibbee, Richard J. [4 ]
Sattar, Syed A. [4 ]
Zamay, Tatiana N. [2 ]
Berezovski, Maxim V. [1 ]
机构
[1] Univ Ottawa, Dept Chem, Ottawa, ON K1N 6N5, Canada
[2] Krasnoyarsk State Med Univ, Inst Mol Med & Pathol Biochem, Krasnoyarsk 660022, Russia
[3] Krasnoyarsk State Med Univ, Dept Microbiol, Krasnoyarsk 660022, Russia
[4] Univ Ottawa, Ctr Res Environm Microbiol, Fac Med, Ottawa, ON K1H 8M5, Canada
基金
加拿大自然科学与工程研究理事会;
关键词
SALMONELLA; DNA; CHOLERAESUIS; GOLD; PCR;
D O I
10.1021/ac302217u
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
The development of an aptamer-based impedimetric sensor for typing of bacteria (AIST-B) is presented. Highly specific DNA aptamers to Salmonella enteritidis were selected via Cell-SELEX technique. Twelve rounds of selection were performed; each comprises a positive selection step against S. enteritidis and a negative selection step against a mixture of related pathogens, including Salmonella typhimurium, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Citrobacter freundii, to ensure the species-specificity of the selected aptamers. After sequencing of the pool showing the highest binding affinity to S. enteritidis, a DNA sequence of high affinity to the bacteria was integrated into an impedimetric sensor via self-assembly onto a gold nanoparticles-modified screen-printed electrode (GNPs-SPCE). Remarkably, this aptasensor is highly selective and can successfully detect S. enteritidis down to 600 CFU mL(-1) (equivalent to 18 CFU in 30 mu L, assay volume) in 10 min and distinguish it from other Salmonella species, including S. typhimurium and S. choleraesuis. This report is envisaged to open a new venue for the aptamer-based typing of a variety of microorganisms using a rapid, economic, and label-free electrochemical platform.
引用
收藏
页码:8114 / 8117
页数:4
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