Development of PCR-based assays for the detection of two human mollicute species, Mycoplasma penetrans and M. hominis

Grau, Odile; Kovacic, Rémi; Griffais, Rémi; Launay, Valérie; Montagnier, Luc

doi:10.1006/mcpr.1994.1019

Cited by 41 publications

(10 citation statements)

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“…PCR assays for M. hominis have used 16S rRNA and ribosomal DNA as the gene targets (100,344). The theoretical advantages of the PCR assay for detection of genital mycoplasmas include the fact that no viable organisms are necessary, its limit of detection is much better than culture, and results can be available in 1 day.…”

Section: Nucleic Acid Amplificationmentioning

confidence: 99%

Mycoplasmas and Ureaplasmas as Neonatal Pathogens

Katz²,

2005

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SUMMARY The genital mycoplasmas represent a complex and unique group of microorganisms that have been associated with a wide array of infectious diseases in adults and infants. The lack of conclusive knowledge regarding the pathogenic potential of Mycoplasma and Ureaplasma spp. in many conditions is due to a general unfamiliarity of physicians and microbiology laboratories with their fastidious growth requirements, leading to difficulty in their detection; their high prevalence in healthy persons; the poor design of research studies attempting to base association with disease on the mere presence of the organisms in the lower urogenital tract; the failure to consider multifactorial aspects of diseases; and considering these genital mycoplasmas only as a last resort. The situation is now changing because of a greater appreciation of the genital mycoplasmas as perinatal pathogens and improvements in laboratory detection, particularly with regard to the development of powerful molecular nucleic acid amplification tests. This review summarizes the epidemiology of genital mycoplasmas as causes of neonatal infections and premature birth; evidence linking ureaplasmas with bronchopulmonary dysplasia; recent changes in the taxonomy of the genus Ureaplasma; the neonatal host response to mycoplasma and ureaplasma infections; advances in laboratory detection, including molecular methods; and therapeutic considerations for treatment of systemic diseases.

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Section: Nucleic Acid Amplificationmentioning

confidence: 99%

Mycoplasmas and Ureaplasmas as Neonatal Pathogens

Katz²,

2005

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“…Although culture is considered to be the reference technique for the detection of M. hominis , it requires specialized media and expertise and does not yield results before 2–5 days [3]. Most PCR techniques detecting M. hominis target the 16S rRNA gene or the 16S‐23S intergenic spacer region by conventional or real‐time assays [4–9]. However, minor sequence variations were observed in the 16S rRNA gene sequences [10] and may lead to a lower clinical sensitivity of these techniques.…”

Section: Introductionmentioning

confidence: 99%

Development of a real-time PCR targeting the yidC gene for the detection of Mycoplasma hominis and comparison with quantitative culture

Férandon

Peuchant

Janis

et al. 2011

Clinical Microbiology and Infection

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Mycoplasma hominis is an opportunistic human mycoplasma species that can be either commensal or pathogenic. Its detection by culture is considered to comprise the reference technique. Previously reported PCR techniques target the 16S rRNA or the gap gene, although sequence variations among clinical isolates may lead to variations in clinical sensitivity. The present study aimed to develop a specific TaqMan quantitative real-time PCR assay, targeting a gene conserved in all M. hominis isolates, and to compare it with quantitative culture. With the knowledge of the M. hominis PG21 genome sequence, the yidC gene, encoding a membrane protein translocase, was chosen as target. Its intraspecies heterogeneity was checked at the nucleotide level using 31 reference or clinical strains. The limit of detection, the analytical specificity and the reproducibility of the assay were assessed. Moreover, PCR and culture results were compared using 153 urogenital specimens. The limit of detection was seven copies/μL. The analytical specificity was 100%, with good inter- and intra-assay reproducibility. Among the 153 urogenital specimens, the yidC PCR and culture allowed detection of 55 and 45 M. hominis-positive samples, respectively. Comparison of the bacterial load among the 45 specimens found to be M. hominis-positive by both techniques revealed a statistically significant association between the quantitative results obtained. In conclusion, we developed a specific, sensitive and reproducible real-time PCR to detect all M. hominis clinical isolates. This PCR was shown to have higher sensitivity than culture, although both methods were correlated for quantification of M. hominis loads in urogenital specimens.

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“…M. penetrans infection was detected in the patient's specimens prior to culture and was confirmed by specific polymerase chain reaction (PCR) (7) (Figure 1A,B). Similar results were obtained by another pair of PCR primers also within the 16S rRNA gene and designed for the specific detection of M. penetrans (data not shown).…”

Section: Casementioning

confidence: 87%

“…A. M. penetrans PCR genomic amplification with the primers MYCPENET-P and MYCPENET-N (7) and analyzed by electrophoresis in 2% agarose gel. Lysates from the following original samples: throat swab (lane 1); tracheal aspirate (lane 2); blood (lane 3); first blood subculture (HF-1 isolate) (lane 4); M. penetrans GTU-54-6A1 (lane 5), showing the amplification product of 407-bp; and negative control (lane 6).…”

Section: Casementioning

confidence: 99%