Abstract
Antibiotic-resistant bacteria present a global threat because the infections they cause are difficult to treat. Therefore, it is highly important to develop advanced methods for the identification of antibiotic resistance gene in the virulent bacteria. Here, we report the development of novel nanoprobes for fluorescence in situ hybridization (FISH) and the application of the nanoprobe to the detection of ampicillin-resistant Escherichia coli. The nanoprobe for FISH was synthesized by the modified sol–gel chemistry and the synthesized nanoprobe provided strong fluorescent signals and pH stability even under natural light condition. For the double-identification of bacteria species and ampicillin-resistance with a single probe in situ, the nanoprobes were conjugated to the two kinds of biotinylated probe DNAs; one for E. coli-species specific gene and the other for a drug-resistant gene. By using the nanoprobe-DNA conjugants, we successfully detected the ampicillin-resistant E. coli through the FISH technique. This result suggests the new insight into light stable FISH application of the nanoprobe for a pathogenic antibiotic-resistance bacterium.
Highlights
Antibiotic-resistant bacteria have been a global issue threatening public health because they are increasingly difficult to treat, leading to higher medical costs, prolonged hospital stays, and increased mortality [1]
In order to conjugate the nanoprobe with single stranded DNAs (ssDNAs), we modified the nanoprobe with APTMS
The resultant amino-modified nanoprobe was reacted with NHS-polyethylene glycol (PEG)-Biotin for 1 h at room temperature
Summary
Antibiotic-resistant bacteria have been a global issue threatening public health because they are increasingly difficult to treat, leading to higher medical costs, prolonged hospital stays, and increased mortality [1]. It is critical to identify the pathogen species and drug-resistant gene accurately in a timely manner for appropriate treatment of antibiotic-resistant bacteria infected patients. For the diagnostics of antibiotic-resistant bacteria, the conventional culture-based plating assay has been widely used. This method requires several days to confirm the growth of the targeted bacterial colony [4]. Since the diagnostic tests are crucial to the management of infectious diseases and combatting the rise in antibiotic resistance, it is urgent to develop the advanced diagnostic methods, which would need to be simple, rapid, accurate, and low cost, for the detection and profiling of antibiotic resistance genes or microorganisms
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