Analysis Identifying Common and Distinct Sequences among Texas Clinical Strains of Mycoplasma genitalium

Abstract

Mycoplasma genitalium is associated with numerous human genitourinary tract maladies, including nonchlamydial, nongonococcal urethritis in men and cervicitis, endometritis, and pelvic inflammatory disease in women (1,4,5,12,19,29,30,35,39). Understanding the epidemiological aspects of infections and developing effective treatment require reliable typing of pathogenic microorganisms. Since M. genitalium is very difficult to culture from clinical specimens (13, 21), classical microbiological typing methods are not readily applicable. Furthermore, serological approaches are not widely used because of cross-reactivity with other mycoplasmas, especially with Mycoplasma pneumoniae (15,26). Therefore, typing of M. genitalium strains relies on DNA sequence data. Recently, sequence variability of the rRNA operon and tandem repeats in the MG309 locus have been evaluated with clinical specimens (28). However, the majority of M. genitalium clinical sequence data available today is based upon the gene mgpB (locus MG191 of sequenced reference strain G37 [11]) encoding the M. genitalium adhesin MgPa (14,17,21). An M. genitaliumspecific diagnostic PCR assay was designed to target the proximal unique region of this gene (using primers MgPa-1 and MgPa-3 [22] and designated here "MgPa-13"). The MgPa-13 region has exhibited sequence stability in persistently infected patients for up to 21 months (17). Despite this intrastrain stability, high levels of sequence variability between clinical isolates were observed (14, 17). Based on this sequence region, transmission of M. genitalium between sexual partners was shown, as was colonization of different anatomical sites of the same patient by identical strains (14).Here, we present analysis of MgPa-13 sequences generated from clinical specimens collected at the Project S.A.F.E. sexually transmitted disease (STD) clinic in San Antonio, TX (36), from recruited female participants and their male partners. Alignment of MgPa-13 sequences corroborated transmission between partners and colonization of different anatomical sites by the same strain. Comparison of newly recovered sequences with all currently available clinical data (14, 17, 21) revealed not only the presence of several common variants but also distinct sequences. Analyses of encoded amino acids uncovered mutations that determine common features among MgPa-13 region variants. MATERIALS AND METHODSOrigin and processing of clinical specimens. A group of minority women (Mexican and African American) and their partners were recruited from public health clinics throughout San Antonio, TX, into a controlled randomized trial of behavioral-cognitive interventions to reduce the recurrence of STDs (36). Women were recruited into the study after being diagnosed with a nonviral STD, specifically infection with Chlamydia trachomatis, Neisseria gonorrheae, Trichomonas vaginalis, or Treponema pallidum. At each clinic visit, urine specimens were obtained from both female and male subjects, and at the same time, vaginal and cervical swab (VC) speci...

Method comparison for molecular typing of French and Tunisian Mycoplasma genitalium-positive specimens

“…In comparison, the mgpB SNP typing resulted in amplification of DNA from 96 % of the specimens, higher than the percentage reported in a recent study using this typing method (38.9 %) (Musatovova & Baseman, 2009). Ma et al (2008) had already applied nested PCR to those specimens for which the initial PCR products were not visible or very faint on agarose gels.…”

“…We used the numbering system developed originally in 2006 by Hjorth et al (2006) with the description of 56 STs, numbered 1-56. In addition to the 56 STs, we successively included the seven new STs described by Ma et al (2008), numbered 57-63, and the 17 new STs described by Musatovova & Baseman (2009), numbered 64-80. In our study, we then described eight new STs of the mgpB gene, numbered 81-88, and deposited them in GenBank under the accession numbers HM565865-HM565872, respectively.…”

“…PCR amplification and sequencing of 281 base pairs (bp) of the MG191 semi-conserved region were performed as previously described using the MgPa-1/MgPa-3 primer set (Jensen et al, 1991). A sequence type (ST) type was assigned to each specimen according to previously described types (Hjorth et al, 2006;Ma et al, 2008;Musatovova & Baseman, 2009). We used the numbering system developed originally in 2006 by Hjorth et al (2006) with the description of 56 STs, numbered 1-56.…”

“…The majority of molecular typing data for M. genitalium clinical specimens is based on the mgpB gene, which encodes the adhesin MgPa (Hjorth et al, 2006;Iverson-Cabral et al, 2006;Musatovova & Baseman, 2009). …”

Sequences of Mycoplasma hominis in Patients with Urinary Tract Infection in a Hospital in Kashan, Iran

“…Microbiol., 10 (6): 260-269, 2015 In a previous study, Kenri et al (2012) reported that Mycoplasma pneumoniae strain 309, a type 2a (subtype 2 variant) strain, has variations in the P1 protein, which is responsible for the attachment of this bacterium to the host cells. Furthermore, Musatovova and Baseman (2009) identified common and distinct sequences among the Texas clinical strains of Mycoplasma genitalium and obtained 18 DNA sequence variants, which were compared with all the other available clinical sequences. A detailed analysis revealed silent mutations of six amino acid residues within the encoded region of the MgPaadhesin in numerous clinical strains.…”