Abstract
BackgroundMycoplasma hominis is an opportunistic human mycoplasma species that can cause various urogenital infections and, less frequently, extragenital infections. The objective of this work was to study the genetic diversity of this species using a molecular typing method based on multiple-locus variable-number tandem repeat (VNTR) analysis (MLVA).ResultsThe genome content of M. hominis PG21 was analysed for tandem repeats (TRs), and five of the 130 TRs identified were selected for use in an MLVA assay. The method was based on GeneScan analysis of VNTR loci using multiplex PCR with fluorescent dyes and resolution by capillary electrophoresis. This approach was used on a collection of 210 urogenital and extragenital French clinical isolates collected between 1987 and 2009. Forty MLVA types were found. The discriminatory index of our MLVA scheme was 0.924. Using this new typing tool, persistent infection was suggested for six patients and new infection for one patient. Furthermore, mother-to-child transmission was confirmed in the two cases studied. Application of MLVA to a wide range of M. hominis isolates revealed high genotypic diversity and no obvious link between the MLVA type and the isolate year of collection, the patient’s age or sex, the anatomical origin of the isolates or resistance to antibiotics was found.ConclusionsOur MLVA scheme highlights the high genetic heterogeneity of the M. hominis species. It seems too discriminatory to be used for large epidemiological studies but has proven its usefulness for molecular studies at the individual level.
Highlights
Mycoplasma hominis is an opportunistic human mycoplasma species that can cause various urogenital infections and, less frequently, extragenital infections
The 167 urogenital clinical isolates were collected at the Bordeaux University Hospital and obtained from i) specimens where M. hominis was present as a commensal, i.e. cervical samples with titres of M. hominis < 104 CCU /ml and male specimens, ii) cervical swabs from patients with titres of M. hominis ≥ 104 CCU /ml without association with BV, iii) cervical swabs from female patients with titres of M. hominis ≥ 104 CCU /ml and suffering from bacterial vaginosis, iv) vaginal swabs from pregnant women with threatened preterm delivery whatever the titre of M. hominis, v) specimens from women presenting upper genital tract infection whatever the titre of M. hominis, these specimens being normally sterile
Identification of variable-number tandem repeat (VNTR) for MLVA typing Among the 130 tandem repeats (TRs) tested, only five were polymorphic with different allele sizes, making them useful for discriminating among types
Summary
Mycoplasma hominis is an opportunistic human mycoplasma species that can cause various urogenital infections and, less frequently, extragenital infections. Pulse-field gel electrophoresis (PFGE) [6,7], restriction fragment length polymorphism (RFLP) analysis [8], amplified fragment length polymorphism (AFLP) [9] and random amplified polymorphic DNA (RADP) [10] have been used to study the genetic diversity of this species. These methods are timeconsuming, require a relatively large amount of biological material, may be difficult to reproduce and standardise between laboratories and generate results that are difficult to interpret
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