Approaches to the detection of whole cells using reflectometric interference spectroscopy

Abstract

AbstractMethods for rapid and reliable detection of clinically relevant bacteria are of highest interest. For such purposes, many analytical methods, e.g. ELISA, have been developed. Also optical biosensors like reflectometric interference spectroscopy (RIfS) are suitable analytical platforms. Here, results are described to capture and detect Legionella pneumophila serogroup 1 on differently functionalized surfaces of RIfS glass chips. As a major problem, availability of suitable antibodies was found. Immobilization of capturing antibodies on the chip surface followed by trapping of L. pneumophila gave only insufficient results. For this approach, the chip surface was partially blocked with bovine serum albumin (BSA) in different ratios between BSA and the Legionella specific antibody in order to obtain a site‐directed immobilization. Further on, antibodies were immobilized by hydrophobic interaction followed by BSA treatment and exposition to L. pneumophila. However, also this approach did not allow sufficient capturing of bacteria cells. It can be assumed that binding forces between the antibody and bacteria are too low. Alternatively, L. pneumophila bacteria were directed immobilized on a hydrophobic chip surface. A sufficient sensor signal for quantification was obtained in a range between 8.25 × 106 and 8.25 × 108 bacteria cells/mL. This approach might open up the possibility for safe identification of L. pneumophila after subsequent addition of specific antibodies.