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The Diagnostic Hybrids' D3 DFA Chlamydiae Culture Confirmation Kit is intended for the qualitative detection of Chlamydiae lipopolysaccharide (LPS) in inoculated cell cultures by immunofluorescence using fluoresceinated monoclonal antibodies (MAbs). Performance has not been established with direct patient specimens.

The Diagnostic Hybrids' D3 DFA Chlamydiae Culture Confirmation Kit includes a Chlamydiae DFA Reagent that contains a blend of two murine MAbs directed against epitopes on the lipopolysaccharide of Chlamydiae. The kit is used for Chlamydiae detection in cell cultures of patient specimens. Kit Components: Chlamydiae DFA Reagent - one dropper bottle containing fluorescein labeled murine MAbs directed against Chlamydiae. The buffered, stabilized, aqueous solution contains Evan's Blue as a counter-stain and 0.1% sodium azide as preservative. Chlamydiae Antigen Control Slides - 10 slides. Five individually packaged control slides with wells containing cell culture-derived Chlamydia trachomatis positive cells and five individually packaged control slides with wells containing cell culture- derived negative cells. Each slide is intended to be stained only one time. PBS Concentrate - a 40X concentrate consisting of 4% sodium azide (after dilution to 1X in water, the concentration of sodium azide in the solution is 0.1%) in PBS Mounting Fluid - an aqueous, buffered, stabilized solution of glycerol and 0.1% sodium azide. The cell cultures to be tested are fixed in acetone. The Chlamydiae DFA Reagent is added to the cells to determine the presence of chlamydial antigens (LPS). After incubating at 35°C to 37°C, the stained cells are rinsed with the diluted PBS Concentrate, a drop of the supplied Mounting Fluid is added and the monolayer is examined for presence of fluorescent inclusions using a fluorescence microscope equipped with the correct filter combination for FITC at a magnification of 100-400X. Chlamydiae infected cells will be stained with bright apple-green fluorescence while uninfected cells will contain no apple-green fluorescence but will fluoresce red by the Evan's Blue counter-stain which is included in the Chlamydiae DFA Reagent.