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Appl. Environ. Microbiol., Feb 1996, 328-331, Vol 62, No. 2
Copyright © 1996, American Society for Microbiology

Application of a Mycoplasma group-specific PCR for monitoring decontamination of Mycoplasma-infected Chlamydia sp. strains

JM Ossewaarde, A de Vries, T Bestebroer and AF Angulo
Research Laboratory for Infectious Diseases, National Institute of Public Health and Environmental Hygiene, Bilthoven, The Netherlands. JM.Ossewaarde@rivm.nl

Mycoplasma contamination of biological materials remains a major problem. Most contaminations are caused by the use of Mycoplasma- contaminated cell lines. We adapted a Mycoplasma group-specific PCR to detect Mycoplasma contamination in cell lines and demonstrate its use in monitoring decontamination procedures with Mycoplasma-contaminated suspensions of Chlamydia spp. Three different methods were investigated: the use of Mycoplasma-specific antiserum in cell culture, physical separation by the combined use of enzymatic treatment and differential centrifugation, and the use of detergents. With these methods only incubation with Triton X-100 resulted in decontamination of Mycoplasma-contaminated suspensions of several laboratory strains of Chlamydia pneumoniae, C. pecorum, and C. trachomatis. Only one C. pneumoniae strain, UZG-1, was sensitive to Triton X-100 treatment. Since 39 of 40 throat swabs from patients with symptoms of an upper respiratory tract infection had positive reactions in the Mycoplasma group-specific PCR, this procedure could also have clinical significance in attempts to propagate C. pneumoniae strains from clinical specimens.

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