| Sumario: | Biofilms are microbial communities attached to a surface, resistant to antimicrobic agents, and causative of medical device-associated and chronic infections. To date, no clinical laboratory test has been identified to assist in diagnosis of biofilm-associated infection. For that reason, this study focused on assay development. Two model systems of bacteria, Staphylococcus aureus and Pseudomonas aeruginosa (Gram positive and Gram negative, respectively) were used for assay development since these organisms frequently are able to produce biofilms and cause antimicrobic-resistant infections. Test format was growth of bacteria in a 96-well microplate using reagents and equipment readily available in clinical laboratories. Procedural steps included inoculation of isolated bacteria into broth and standardization of bacterial number using spectrophotometer or McFarland density standard comparison. Bacteria were dispensed into microplate wells, incubated to allow for biofilm formation; biofilm was then stained, solubilized and quantitated by spectrophotometry. Results indicated that bacterial biofilm is quantifiable using the assay herein defined. In addition, S. aureus and P. aeruginosa are able to produce large amounts of biofilm; and MRSA produces more biofilm than MSSA.
|
|---|