This preparation has tested positive for Mycoplasma contamination.
Note that activities with high potential for aerosol production require BSL 3 facilities and practices.
Forms intracellular inclusions which react with specific anti-C. pneumoniae monoclonal antibodies and monoclonal antibodies directed against the chlamydia LPS.
Inclusions will not stain with iodine or monoclonal antibodies against the MOMP of C. trachomatis. Infection is enhanced by centrifugation and use of cycloheximide (1 to 2 mcg/ml) in medium.
Suggested protocol for propagation: Add glass beads and vortex preparation to disrupt cells. Infect monolayer with disrupted material. Centrifuge at 3000 x rpm (750 x g) for 1 hour.
Feed with fresh growth medium containing FBS prescreened for Chlamydia antibodies and 1-2 ug/mL cycloheximide. Incubate at 37°C for 72 hours.
-70°C or colder
Infection is enhanced by centrifugation and use of cycloheximide (1 to 2 mcg/ml) in medium.
Effect on Host
Intracellular inclusion bodies visualized by fluorescent staining with genus or species specific conjugated antibodies