As specimens used in this study are part of routine patient management without any additional sampling, and since patients provided no objection for their samples to be used, the article L1211-2 of the French code of Public Health states that this study did not need to be examined by the ethical committee “Comité de Protection des Personnes” and that patient’s informed consent was not required. Bacterial strains, culture and DNA preparation The PG21 (ATCC 23114), M132 (ATCC 43521) and H34 (ATCC 15056) M. hominis reference strains and 207 French clinical isolates collected between 1987 and 2009 were used in this study (Additional file 1: Table
S1). The 167 urogenital clinical CP673451 purchase isolates were collected at the buy GSK2126458 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. Thirty-four isolates obtained from extragenital specimens and collected this website at hospitals from 10 different French cities were also tested. Finally, we genotyped six isolates obtained from two mother-neonate pairs. Among these 210 isolates, concomitant and sequential isolates were obtained for one and seven patients, respectively. Antibiotic susceptibility testing, realised when M. hominis was in a pathogenic situation, showed that 66 urogenital isolates were resistant to tetracyclines, seven extragenital isolates were resistant to ofloxacin, two urogenital isolates were resistant to both tetracyclines
and ofloxacin and 91 isolates presented a wild-type profile. The growth conditions used for the M. hominis isolates have been described previously [21]. The DNA was extracted using the MagNA Pure LC DNA isolation kit I (Roche, Meylan, France) according to the manufacturer’s instructions. MLVA analysis https://www.selleckchem.com/products/Roscovitine.html Tandem repeat (TR) sequences were identified in the M. hominis PG21 genome [20] using the Tandem Repeats Finder programme (http://tandem.bu.edu/trf/trf.html) [22]. Loci were chosen if they had >80% matches between the DNA sequences of the repeat units. A total of 130 TRs were selected and designated by the letters Mho followed by a number corresponding to the order in which the TR was detected. To screen for variability in the number of TRs, PCR primers targeting the regions flanking TR loci were designed and tested on a set of 12 M.