Development of PCR-based assays for the detection of two human mollicute species, Mycoplasma penetrans and M. hominis.
Recently, a 16S rDNA-based polymerase chain reaction (PCR) assay was developed for the selective and sensitive detection of Mycoplasma pirum. In this study, the same procedure was used in order to selectively detect by PCR two human mycoplasmas, M. hominis and M. penetrans, with a high level of sensitivity even in a context of human DNA. For each assay, the specificity was verified by testing DNA from other mollicute species (including those closely related to the corresponding mycoplasma), from bacteria phylogenetically close to mollicutes, from Escherichia coli and from human peripheral blood mononuclear cells (PBMCs). Each assay proved to be highly sensitive since it reliably detected 10 DNA molecules, even in a context of human DNA. The results of this study demonstrate the suitability of our procedure using primers which were designed for the PCR detection of human mollicutes with a high specificity and a low and reproducible threshold of sensitivity.
Grau
O
O
Unité d'Oncologie Virale, Institut Pasteur, Paris, France.
Kovacic
R
R
Griffais
R
R
Launay
V
V
Montagnier
L
L
D001483:Base Sequence; D015139:Blotting, Southern; D004269:DNA, Bacterial / Q000032:analysis* / Q000097:blood / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D009174:Mycoplasma / Q000235:genetics* / Q000302:isolation & purification; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity
D004269:DNA, Bacterial; D015345:Oligonucleotide Probes