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Development of a Capillary Biosensor for Helicobacter hepaticus

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Abstract:

Helicobacter hepaticus is a bacterium that causes chronic active hepatitis in mice. Early detection is necessary to prevent the spread of infection and the invalidation of research studies involving infected mice. Current sensing mechanisms are time consuming, insensitive, or are not suitable for screening. The objective of this study was to develop a fast, accurate capillary biosensor for the detection of H. hepaticus. A capillary biosensor platform was designed and fabricated, and a competitive immunoassay was adapted for use in this platform. H. hepaticus was immobilized to the inner wall of the capillary. A fixed number of anti-H. hepaticus antibodies conjugated to Alexa Fluor 546 was added to samples containing H. hepaticus and served as a fluorescent tracer element. Sample H. hepaticus bound to the antibodies in solution, thereby preventing the antibodies from attaching to the immobilized H. hepaticus in the capillary waveguides. An increase in sample H. hepaticus decreased the number of antibodies bound to the capillary wall. The H. hepaticus immobilized capillary waveguides were exposed to different concentrations of the labeled antibody/H. hepaticus solutions. The results showed that the capillary biosensors had a 50 ng/ml (0.75 ng) limit of detection or less, but greater than 5 ng/ml, in an assay time of approximately one hour. Additionally, the capillary biosensors were exposed to mice fecal matter and the results indicated that the biosensor could distinguish between positive and negative samples. The biosensor was shown to reliably detect low levels of H. hepaticus.

Keywords: CAPILLARY; FLUORESCENCE; HELICOBACTER HEPATICUS; IMMUNOSENSOR

Document Type: Research Article

DOI: http://dx.doi.org/10.1166/sl.2007.211

Publication date: June 1, 2007

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