Open AccessDifferentiation of borreliacidal activity caused by immune serum or antimicrobial agents by flow cytometry
Author(s) -
Y F Liu,
L. C. L. Lim,
Kathleen Schell,
Steven D. Lovrich,
Steven M. Callister,
Ronald F. Schell
Publication year - 1994
Publication title -
clinical and diagnostic laboratory immunology
Language(s) - English
Resource type - Journals
eISSN - 1098-6588
pISSN - 1071-412X
DOI - 10.1128/cdli.1.2.145-149.1994
Subject(s) - antimicrobial , flow cytometry , immune system , acridine orange , penicillin , microbiology and biotechnology , antibody , fluorescein isothiocyanate , biology , antibody dependent cell mediated cytotoxicity , chemistry , immunology , antibiotics , monoclonal antibody , biochemistry , apoptosis , fluorescence , physics , quantum mechanics
We demonstrated that borreliacidal activity caused by immune serum and complement can easily be differentiated by flow cytometry from killing activity caused by antimicrobial agents that are commonly used for the treatment of Lyme disease. Assay suspensions containing normal or immune serum were incubated with Borrelia burgdorferi in the presence or absence of ceftriaxone, doxycycline, penicillin, and phosphomycin for 2, 8, 16, and 24 h. Samples containing killing activity were identified by using flow cytometry and acridine orange. In 30 min, the effects of immune serum and complement were easily distinguished from the killing of spirochetes by antimicrobial agents by adding fluorescein isothiocyanate-conjugated goat anti-hamster immunoglobulin. This simple procedure greatly enhanced the usefulness of the borreliacidal assay by eliminating a major source of false-positive reactions.