Diversity of Mycoplasma hominis clinical isolates from Bordeaux, France, as assessed by multiple-locus variable-number tandem repeat analysis

BMC Microbiology

Cécile Bébéar¹  Hélène Renaudin¹  Olivia Peuchant¹  Cyril Férandon² 

[1] CHU de Bordeaux, Laboratoire de Bactériologie, 33076, Bordeaux, France;INRA, USC Mycoplasmal and Chlamydial Infections in Humans, 33076, Bordeaux, France

关键词: Genotyping;    MLVA;    Mycoplasma hominis;   

Others  :  1143687
DOI  :  10.1186/1471-2180-13-120

 received in 2012-11-27, accepted in 2013-05-24,  发布年份 2013

【 摘 要 】

Background

Mycoplasma 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).

Results

The 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.

Conclusions

Our 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.

【 授权许可】

   
2013 Férandon et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

【 参考文献 】

• [1]Waites KB, Schelonka RL, Xiao L, Grigsby PL, Novy MJ: Congenital and opportunistic infections: Ureaplasma species and Mycoplasma hominis. Semin Fetal Neonatal Med 2009, 14(4):190-199.
• [2]Lund KJ, Williamson C, Mc Gregor JA, Parsons A: Uterine transport of ultrasound contrast medium in non-pregnant women: a key to understanding bacterial vaginosis (BV) associated endometritis and salpingitis. Int J Gynecol Obstet 1999, 67(Suppl1):S52.
• [3]Taylor-Robinson D, Jensen JS, Svenstrup H, Stacey CM: Difficulties experienced in defining the microbial cause of pelvic inflammatory disease. Int J STD AIDS 2012, 23(1):18-24.
• [4]Mardh PA: Mycoplasmal PID: a review of natural and experimental infections. Yale J Biol Med 1983, 56(5–6):529-536.
• [5]Waites KB, Talkington DF: New developments in human diseases due to mycoplasmas. In Mycoplasmas molecular biology pathogenicity and strategies for control. Edited by Blanchard A, Browning GF. Norfolk: Horizon Bioscience; 2005:289-354.
• [6]Ladefoged SA, Christiansen G: Physical and genetic mapping of the genomes of five Mycoplasma hominis strains by pulsed-field gel electrophoresis. J Bacteriol 1992, 174(7):2199-2207.
• [7]Jensen LT, Thorsen P, Moller B, Birkelund S, Christiansen G: Antigenic and genomic homogeneity of successive Mycoplasma hominis isolates. J Med Microbiol 1998, 47(8):659-666.
• [8]Busch U, Nitschko H, Pfaff F, Henrich B, Heesemann J, Abele-Horn M: Molecular comparison of Mycoplasma hominis strains isolated from colonized women and women with various urogenital infections. Zentralbl Bakteriol 2000, 289(8):879-888.
• [9]Kokotovic B, Friis NF, Jensen JS, Ahrens P: Amplified-fragment length polymorphism fingerprinting of Mycoplasma species. J Clin Microbiol 1999, 37(10):3300-3307.
• [10]Soroka AE, Momynaliev KT, Taraskina AM, Savicheva AM, Govorun VM: Genetic heterogeneity of Mycoplasma hominis clinical isolates detected during observation of patients with recurrent urogenital inflammation. Bull Exp Biol Med 2001, 132(1):663-665.
• [11]Boesen T, Emmersen J, Baczynska A, Birkelund S, Christiansen G: The vaa locus of Mycoplasma hominis contains a divergent genetic islet encoding a putative membrane protein. BMC Microbiol 2004, 4:37. BioMed Central Full Text
• [12]Mardassi BB, Ayari H, Bejaoui-Khiari A, Mlik B, Moalla I, Amouna F: Genetic variability of the P120’ surface protein gene of Mycoplasma hominis isolates recovered from Tunisian patients with uro-genital and infertility disorders. BMC Infect Dis 2007, 7:142. BioMed Central Full Text
• [13]Mygind T, Birkelund S, Christiansen G: Characterization of the variability of a 75-kDa membrane protein in Mycoplasma hominis. FEMS Microbiol Lett 2000, 190(1):167-176.
• [14]Sogaard IZ, Boesen T, Mygind T, Melkova R, Birkelund S, Christiansen G, Schierup MH: Recombination in Mycoplasma hominis. Infect Genet Evol 2002, 1(4):277-285.
• [15]Cazanave C, Charron A, Renaudin H, Bébéar C: Method comparison for molecular typing of French and Tunisian Mycoplasma genitalium-positive specimens. J Med Microbiol 2012, 61(Pt 4):500-506.
• [16]Dégrange S, Cazanave C, Charron A, Renaudin H, Bébéar C, Bébéar CM: Development of multiple-locus variable-number tandem-repeat analysis for molecular typing of Mycoplasma pneumoniae. J Clin Microbiol 2009, 47(4):914-923.
• [17]McAuliffe L, Ayling RD, Nicholas RA: Identification and characterization of variable-number tandem-repeat markers for the molecular epidemiological analysis of Mycoplasma mycoides subspecies mycoides SC. FEMS Microbiol Lett 2007, 276(2):181-188.
• [18]Pinho L, Thompson G, Rosenbusch R, Carvalheira J: Genotyping of Mycoplasma bovis isolates using multiple-locus variable-number tandem-repeat analysis. J Microbiol Methods 2012, 88(3):377-385.
• [19]Vranckx K, Maes D, Calus D, Villarreal I, Pasmans F, Haesebrouck F: Multiple-locus variable-number tandem-repeat analysis is a suitable tool for differentiation of Mycoplasma hyopneumoniae strains without cultivation. J Clin Microbiol 2011, 49(5):2020-2023.
• [20]Pereyre S, Sirand-Pugnet P, Beven L, Charron A, Renaudin H, Barré A, Avenaud P, Jacob D, Couloux A, Barbe V, et al.: Life on arginine for Mycoplasma hominis: clues from its minimal genome and comparison with other human urogenital mycoplasmas. PLoS Genet 2009, 5(10):e1000677.
• [21]Waites KB, Bébéar C, Robertson JA, Talkington DF, Kenny GE: Cumitech 34, Laboratory diagnosis of mycoplasmal infections. Coordinating edition. Washington, DC: F. S. Nolte. American Society for Microbiology; 2001.
• [22]Benson G: Tandem repeats finder: a program to analyze DNA sequences. Nucleic Acids Res 1999, 27(2):573-580.
• [23]Hunter PR, Gaston MA: Numerical index of the discriminatory ability of typing systems: an application of Simpson’s index of diversity. J Clin Microbiol 1988, 26(11):2465-2466.
• [24]Pereyre S, Gonzalez P, De Barbeyrac B, Darnige A, Renaudin H, Charron A, Raherison S, Bébéar C, Bébéar CM: Mutations in 23S rRNA account for intrinsic resistance to macrolides in Mycoplasma hominis and Mycoplasma fermentans and for acquired resistance to macrolides in M. hominis. Antimicrob Agents Chemother 2002, 46(10):3142-3150.
• [25]Grattard F, Soleihac B, De Barbeyrac B, Bébéar C, Seffert P, Pozzetto B: Epidemiologic and molecular investigations of genital mycoplasmas from women and neonates at delivery. Pediatr Infect Dis J 1995, 14(10):853-858.
• [26]Bébéar CM, Kempf I: Antimicrobial therapy and antimicrobial resistance. Wymondham, United Kingdom: Mycoplasmas: pathogenesis, molecular biology, and emerging strategies for control Horizon Bioscience; 2005:535-568. [A Blanchard and GF Browning (ed)]
• [27]Dégrange S, Renaudin H, Charron A, Bébéar C, Bébéar CM: Tetracycline resistance in Ureaplasma spp. and Mycoplasma hominis: prevalence in Bordeaux, France, from, to 2002 and description of two tet(M)-positive isolates of M. hominis susceptible to tetracyclines. Antimicrob Agents Chemother 1999, 52(2):742-744.
• [28]Mardassi BB, Aissani N, Moalla I, Dhahri D, Dridi A, Mlik B: Evidence for the predominance of a single tet(M) gene sequence type in tetracycline-resistant Ureaplasma parvum and Mycoplasma hominis isolates from Tunisian patients. J Med Microbiol 2012, 61(Pt 9):1254-1261.