Application of a Mycoplasma group-specific PCR for monitoring decontamination of Mycoplasma-infected Chlamydia sp. strains.
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.
Ossewaarde
J M
JM
Research Laboratory for Infectious Diseases, National Institute of Public Health and Environmental Hygiene, Bilthoven, The Netherlands. JM.Ossewaarde@rivm.nl
de Vries
A
A
Bestebroer
T
T
Angulo
A F
AF
D001483:Base Sequence; D002689:Chlamydia / Q000235:genetics* / Q000254:growth & development / Q000302:isolation & purification; D002692:Chlamydia trachomatis / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D016993:Chlamydophila pneumoniae / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D002691:Chlamydophila psittaci / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D017931:DNA Primers / Q000235:genetics; D015342:DNA Probes / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D005069:Evaluation Studies as Topic; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D009174:Mycoplasma / Q000235:genetics* / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers; D015342:DNA Probes; D004269:DNA, Bacterial