Biofilms are communities of bacteria, often containing multiple species, and can be found attached to a wide variety of surfaces including soft tissue, bone, catheters, and metals, making the human body and indwelling medical implants attractive targets for infection. Colonization of these materials leads to severe complications such as endocarditis, osteomyelitis, pneumonia, and medical-device compromise or failure. The time between an established biofilm infection and the diagnosis of such infection is determinant of how successful the treatment outcome will be. Studies have shown that traditional culturing methods used in clinical laboratories often fail to detect the presence of a biofilm infection. We have developed a non-culturing diagnostic test that is specific for biofilm infections. By implementing immunogenic and biofilm-specific proteins into a lateral flow assay system we are able to identify biofilm-specific antibodies in serum samples. The antibodies form a complex between colloidal gold and protein, generating results in less than 10 minutes. Blood samples from rabbits with chronic osteomyelitis and rabbits vaccinated with purified biofilm-specific proteins were used to determine specificity of the lateral flow assay. Additionally, we have linked biofilm-specific antibodies to paramagnetic nanoparticle probes that can be used to view a biofilm infection in vivo without removal of the colonized target. These assay systems are designed to produce rapid and sensitive results in hopes of optimizing patient outcome.