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Background: Biofilms are complicated structures used by pathogens to survive in vivo. The immunocompetent host cannot eliminate the pathogens in biofilms. Such polymicrobial infections are highly difficult to treat due to the resistance offered by the bacteria thriving in a symbiotic relationship enabling suitable growth conditions for their survival. In an attempt to recognize the organisms that are found in biofilms, we have established a SPR-based method to distinguish pathogens by their interaction to specific antibodies that were developed in vitro. Methods Blood cells were separated by gradient centrifugation. B-cells were cultured with and without bacteria in L-15 cell culture medium and incubated at 37ºC. Blood cell components were identified by Flow Cytometry. Antibodies were produced in vitro and were analysed by SDS-PAGE. •Immobilization of Antibodies on CM5 sensor chip of Biacore 1000. Bands were obtained at 25 and 50KDa for light and heavy chains of antibodies respectively. Results : Aim The study aims to use SPR system to detect the most common bacteria found in biofilms to treat infections. The medium contained antibodies against incubated bacteria that was immobilised on chip in Biacore system. B cells that were incubated with bacteria were developed to mature B cells(CD19+ly) as shown by flow cytometry analysis. Ulcer secretion from healthy/patients were analysed by Biacore and the results correlated well to the culture results. Conclusion In the present study an In vitro model of antibody production against bacteria is presented. Detection of bacteria by specific antibody interaction in SPR system might be used to identify the bacterial flora in secretions especially when they are infected by several bacterial strains or when the culture and PCR analyses are negative . Detection of bacterial flora in biological secretions using antibodies developed in vitro and immobilised in a surface plasmon resonance system Contact Info: Nakka Sravya Sowdamini [email protected] Acknowledgements I would like to thank my supervisor and Johanna Lonn for their guidance throughout the study. I am also grateful to peas Institut for financially supporting my study. Master Thesis by Nakka Sravya Sowdamini International Masters Program, 2011 Molecular Genetics and Physiology Supervisor: Dr Fariba Nayeri