DNA aptamers were developed against lipopolysaccharide (LPS) from E. coli O111:B4 and shown to bind both LPS and E. coli by a colorimetric enzyme-based microplate assay. The polyclonal aptamers were coupled to human C1qrs protein either directly using a bifunctional linker or indirectly using biotinylated aptamers and a streptavidin-C1qrs complex. Both systems significantly reduced colony counts when applied to E. coli O111:B4 and K12 strains across a series of 10x dilutions of the bacteria in the presence of human serum; it was diluted 1: 10(3) in order to avoid significant bacterial lysis by the competing alternate pathway of complement activation. A number of candidate DNA aptamer sequences were cloned and sequenced from the anti-LPS aptamer library for future screening of antibacterial or "antibiotic" potential and to aid in eventual development of an alternative therapy for antibiotic-resistant bacterial infections.

• MeSH
• antibakteriální látky chemie   imunologie   farmakologie   MeSH
• aptamerová technika SELEX MeSH
• aptamery nukleotidové chemie   genetika   imunologie   farmakologie   MeSH
• Escherichia coli účinky léků   imunologie   MeSH
• infekce vyvolané Escherichia coli farmakoterapie   imunologie   MeSH
• komplement C1 chemie   imunologie   MeSH
• lidé MeSH
• lipopolysacharidy chemie   imunologie   MeSH
• molekulární sekvence - údaje MeSH
• sekvence nukleotidů MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH
• Názvy látek
• antibakteriální látky MeSH
• aptamery nukleotidové MeSH
• komplement C1 MeSH
• lipopolysacharidy MeSH

A novel immunosensing device based on a piezoelectric sensor for direct detection of the biological warfare agent Francisella tularensis was developed. This sensor includes mouse polyclonal antibody immobilized in a layer of protein A covalently linked to the gold electrode of the sensor. The immunosensor is able to detect F. tularensis with the limit of detection 10(5) CFU/mL with a typical measuring cycle > 5 min. The sensor was successfully evaluated for rapid detection of F. tularensis spikes in drinking water and milk; no deterioration of sensitivity in comparison with buffer solutions was observed. The proposed concept of a rapid measurement of microbial agents seems to be promising for evaluation of samples after short pre-cultivation enrichment.

• MeSH
• biologické bojové látky MeSH
• biosenzitivní techniky metody   MeSH
• Francisella tularensis izolace a purifikace   MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• protilátky bakteriální chemie   MeSH
• tularemie mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• biologické bojové látky MeSH
• protilátky bakteriální MeSH