Establishment of a Visual and Rapid Detection Method for Enterotoxigenic Escherichia coli K88
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Abstract:
A method that could rapidly, sensitively, specifically, and visually detect E. coli K88 was established based on the aptamer recognition of the E. coli K88 fimbriae protein, combined with gold nanoparticles (AuNPs) labeling and silver enhancement signal amplification. Biotinylated aptamer 1 that could specifically bind to E. coli K88 was incubated with target bacterium E. coli K88 and the AuNPs-aptamer 2 conjugates, to form a sandwich-type aptamer 1-E. coli K88-aptamer 2-AuNPs complex. Then, the sandwich-type complex was immobilized onto the surface of a 96-microwell plate modified with streptavidin through the binding of biotin and streptavidin. Finally, the responding signal was amplified using silver enhancement. After the optimization of the detection conditions, this method could detect E. coli K88 specifically and quantitatively. When the E. coli K88 concentration was in the range of 1.0×101~1.0×105 cfu/well, the coefficient of determination (R2) for quantitative linear curve fitting was 0.9903, the detection sensitivity could reach 10 cfu/well, and negative results were obtained in the detection of other non-target bacterial strains. This paper provides a basis for the visual detection for E. coli K88 in clinical samples.
