title: Sensitivity and specificity of five different mycoplasma detection assays.
abstract: The sensitivity and specificity of five different mycoplasma detection tests were evaluated in comparison with the classical microbiological culture assay on agar plates as the reference method: direct fluorochrome DNA staining (direct DAPI), DNA staining of an indicator cell line (indirect DAPI), RNA hybridization with a cDNA specific for ribosomal mycoplasmal RNA, an enzyme-linked immunosorbent assay (ELISA) with mycoplasma-specific antibodies, and a biochemical cytotoxicity assay (6-MPDR). A large panel of continuous cell lines (20 adherent and 233 suspension cell lines, most of the latter were human leukemia-lymphoma cell lines) were analyzed for infection with mycoplasma. The results of the comparative analysis for sensitivity and specificity of the various tests were as follows: 100% and 100% for the indirect DAPI, 100% and 98% for the RNA hybridization assay, 87% and 94% for the direct DAPI, 72% and 100% for the ELISA, 75% and 90% for the biochemical 6-MPDR assay. Each of these approaches has both advantages and disadvantages with regard to cost, time, reliability, specificity, and sensitivity. The best compromise for routine mycoplasma testing is a combination of several techniques (e.g. direct culture on agar, RNA hybridization, and direct or indirect DAPI).
authors.affiliation: German Collection of Microorganisms and Cell Cultures, Human and Animal Cell Cultures Collection, Braunschweig.
authors.forename: C C
authors.initials: CC
authors.lastname: Uphoff
authors.forename: S
authors.initials: S
authors.lastname: Brauer
authors.forename: D
authors.initials: D
authors.lastname: Grunicke
authors.forename: S M
authors.initials: SM
authors.lastname: Gignac
authors.forename: R A
authors.initials: RA
authors.lastname: MacLeod
authors.forename: H
authors.initials: H
authors.lastname: Quentmeier
authors.forename: K
authors.initials: K
authors.lastname: Steube
authors.forename: M
authors.initials: M
authors.lastname: Tümmler
authors.forename: M
authors.initials: M
authors.lastname: Voges
authors.forename: B
authors.initials: B
authors.lastname: Wagner
mesh_terms: D000818:Animals; D001431:Bacteriological Techniques; D004269:DNA, Bacterial / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D051379:Mice; D009174:Mycoplasma / Q000235:genetics / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization; D011237:Predictive Value of Tests; D011684:Purine Nucleosides; D012329:RNA, Bacterial / Q000032:analysis; D012335:RNA, Ribosomal / Q000032:analysis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D004269:DNA, Bacterial; D011684:Purine Nucleosides; D012329:RNA, Bacterial; D012335:RNA, Ribosomal; C032885:6-methylpurine 2'-deoxyriboside