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

Abstract

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.

Development and Evaluation of an Immuno-Capture Enzyme-Linked Immunosorbent Assay to Quantify the Mycoplasma capricolum subsp. capripneumoniae (Mccp) Protein in Contagious Caprine Pleuropneumonia (CCPP) Vaccine

“…e first PCR reactions were done with DNA using the Mycoplasma genus-specific primers: upstream primer GPO-3, 5′-gggagcaaacaggattagataccct-3′ and downstream primer MGSO, 5′-tgcaccatctgtcactctgttaacctc-3′. ese were used to amplify a 280 bp fragment [24]. In the second PCR reaction, the Mccp specific primers Mccp MCCPF 5′-atcatttttaatcccttcaag-3′ and MCCPR 5′-tactatgagtaattataatatatgcaa-3′ were used for amplification of a 316 bp fragment sequence [25,26].…”

Centrosome abnormalities during aChlamydia trachomatisinfection are caused by dysregulation of the normal duplication pathway

“…Chlamydia trachomatis EBs were tested for Mycoplasma contaminations using Mycoplasma group-specific PCR as described (Ossewaarde et al, 1996).…”

A Chlamydia pneumoniae infection model using established human lymphocyte cell lines

“…pneumoniae TW183 strain was kindly provided by G. Byrne, University of Wisconsin, Madison, WI, USA. The bacterial cultures were con¢rmed Mycoplasma-free by PCR, as described previously [16]. The bacteria were propagated in the HEp-2 cell culture system according to methods described previously [17].…”