Rapid Identification of Bacterial Virulence Factors

Rapid Virulence Factor Discovery System (RVFDS) is virulence factor-discovery platform in which all of the genes of a pathogen are expressed generating a random expression library (REL). The REL is then screened for the presence of potential virulence factors with an array of pull-down assays that employ components of the host"s extracellular matrix, mucus, and basement membrane as bait. The host bait proteins interact with and capture pathogen prey proteins that may play a role in the pathogenic process. The potential virulence factors are then identified by mass spectrometry and proteomic methodologies. RVFDS can also define the immunome or complete repertoire of immunogenic proteins of a pathogen using antibodies from convalescent patients as bait in pull-down assays. Presently investigations into the host-pathogen relationship are severely hampered by the fact that the pathogen is grown under laboratory conditions that simulate the host environment, and thus, many of the virulence factors that are expressed in response to the true host environment are overlooked. RVFDS circumvents this overwhelming challenge because virtually all of the open reading frames of a pathogen are expressed and are easily and rapidly identified.