This new immunostaining process, used with in vitro cultivation, provides for precise identification of cultured organisms

Lyme disease in humans is caused by Borrelia burgdorferi sensu lato, organisms that
are known to attach to collagen in the body. The use of a matrix containing collagen
as a part of the culture composition has given us the ability to cultivate Borrelia
burgdorferi, B. afzelii, and B. garinii from the bloodstream using artificial media. The
cultured Borrelia is identified by immunostaining using polyclonal and monoclonal
antibodies. In order to distinctly differentiate the organisms from the collagen of this
matrix that could be observed as background in the staining process, we developed
an immunostaining procedure using polyclonal and monoclonal antibodies in
combination with rhodamine fibronectin. The culture samples from both control
organisms and patient samples were tested using the new immunostaining protocol.
Results showed clear delineation of organisms compared to the collagen pieces
gathered in the harvesting process. This new immunostaining process, used with in
vitro cultivation, provides for precise identification of cultured organisms.
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