Genotyping analysis of Mycoplasma pneumoniae clinical strains in Japan between 1995 and 2005: type shift phenomenon of M. pneumoniae clinical strains

Kenri, Tsuyoshi; Okazaki, Norio; Yamazaki, Tsutomu; Narita, Mitsuo; Izumikawa, Kinich; Matsuoka, Masao; Suzuki, Satowa; Horino, Atsuko; Sasaki, Tsuguo

doi:10.1099/jmm.0.47634-0

Cited by 87 publications

(131 citation statements)

References 25 publications

Supporting

Mentioning

114

Contrasting

Unclassified

Order By: Relevance

“…The existence of these two types of the P1 gene (type 1 and type 2) was shown to be an excellent marker for numerous other variations within their genomes, some of which likely lead to virulence differences observed among strains (8). It is possible that well-documented shifts of clinical M. pneumoniae isolates in different locations over time could be a result of type-associated differences (23). One hypothesis speculates that type-specific antibodies render protection against the causative type strain, leading to replacement by the other strain type.…”

Section: Discussionmentioning

confidence: 99%

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

Musatovova

Baseman

2009

J Clin Microbiol

View full text Add to dashboard Cite

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...

show abstract

Section: Discussionmentioning

confidence: 99%

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

Musatovova

Baseman

2009

J Clin Microbiol

View full text Add to dashboard Cite

show abstract

“…Furthermore, in recent years, three variants of M. pneumoniae showing sequence variations in one of these two repetitive elements were described (4, 6, 11). Variant 2a strains were reported with increasing numbers in Japan (12) and Europe (6), but only one isolate has been characterized as variant 1. Variant 2b isolates were detected in limited number with a molecular subtyping method in Europe only (6); therefore, DNA samples from cultures of this genotype could not be included in the present study.…”

mentioning

confidence: 99%

Comparison of Commercial and In-House Real-Time PCR Assays Used for Detection of Mycoplasma pneumoniae

Dumke

Jacobs

2009

J Clin Microbiol

View full text Add to dashboard Cite

We tested two commercial and three in-house PCR assays under standardized conditions to detect Mycoplasma pneumoniae. All five procedures were able to demonstrate M. pneumoniae DNA in a concentration comparable to 1 CFU/l, but the mean crossing points resulted in differences in the concentration of the genome copies of a factor of 20.Recent studies have shown that the cell wall-less mollicute species Mycoplasma pneumoniae is the causative agent of about 5 to 20% of all cases of community-acquired pneumonia in humans (2). In closed populations like those in army bases or in periods showing epidemic peaks of M. pneumoniae infections (occurring every 3 to 7 years) (8, 14), the incidence of infections might increase up to 50% (2). The primarily respiratory tract infection can be followed by extrapulmonary complications (22). The specific antibiotic treatment of the disease, restricted to macrolides, tetracyclines, or fluoroquinolones, accounts for the search for sensitive, specific, and fast methods for the detection of M. pneumoniae infections in routine bacteriological practice. Cultivation is time consuming (up to 3 weeks), insensitive, and requires confirmation that the colonies grown are M. pneumoniae bacteria. Serological methods, such as the complement fixation test, enzyme-linked immunosorbent assay, and Western blot, are commonly used in the clinical laboratory but have practical limitations. The age-and timedependent formation of specific immunoglobulin A and immunoglobulin M antibodies, the persistence of detectable antibody levels resulting from previous contacts with M. pneumoniae, and the variable specificity and sensitivity of the test kits used influence the significance of the results of serodiagnosis (2). Therefore, PCR approaches extended the spectrum of available routine methods. Recent results reported the superiority of PCR over serology for the confirmation of a M. pneumoniae infection in the clinically important first week after the onset of pneumonia symptoms (17).In recent years, different PCR systems targeting a wide range of genes of M. pneumoniae (e.g., P1 adhesin, 16S rRNA, and ATPase operon gene) have been developed (16). Real-time PCR especially is characterized by rapidity, practicability, reduced risk of contamination, and high specificity and sensitivity of detection (3,7,9,13,15,19,23,24,25). However, the increasing number of real-time PCR approaches stresses the need for comparative validation of the performance of the different test procedures. Up to now, studies dealing with the evaluation of more than a single quantitative PCR system to detect M. pneumoniae have been very rare (7, 25) and have not included commercial test kits. To our knowledge, commercial quantitative PCR systems for the detection of M. pneumoniae are not available in the United States (2), whereas kits from different manufacturers are widely used in Europe. The aim of the present study was to investigate the sensitivities of different commercial and in-house real-time PCR approaches for the detection of M. p...

show abstract

“…Sixty of 201 M. pneumoniae clinical isolates were genotyped by full-length sequencing of the p1 gene in our previous report (8); the other 141 isolates were genotyped by a previously reported method (15,16). Results indicated that 184 (91.5%) isolates were type 1, 2 (1.0%) isolates were variant 2a, 15 (7.5%) isolates were variant 2c, and no type 2 isolates were detected.…”

mentioning

confidence: 99%

Multiple-Locus Variable-Number Tandem-Repeat Analysis of 201 Mycoplasma pneumoniae Isolates from Beijing, China, from 2008 to 2011

et al. 2013

View full text Add to dashboard Cite

A total of 201 Mycoplasma pneumoniae clinical isolates from Beijing, China, isolated from 2008 to 2011, were clustered into 16 multiple-locus variable-number tandem-repeat analysis (MLVA) types, of which 6 new MLVA types have never been reported previously. Type 1 isolates based on p1 gene genotyping were mainly MLVA types E, J, P, U, and X. There was no correlation between macrolideresistant Mycoplasma pneumoniae and their MLVA type. Mycoplasma pneumoniae is an important pathogen for human respiratory tract infection, especially for communityacquired pneumonia (CAP), of which M. pneumoniae infection accounts for a ratio of 10% to 30% (1, 2). The genotyping of clinical isolates is an important means for understanding the epidemiology of M. pneumoniae and the analysis of its breakout. Except for fewer adhesion-related protein-coding genes, the genomic sequences of M. pneumoniae are highly homogeneous, so the previously reported genotyping target site of M. pneumoniae is mainly a p1 adhesion protein gene. Commonly used PCR-restriction fragment length polymorphism (PCR-RFLP) technology primarily classified M. pneumoniae into type 1 and type 2 (3, 4). With the in-depth study of the RepMp2/3 and RepMp4 repeat sequence within the p1 gene, more variants of type 1 and 2 isolates were found by molecular subtyping and sequencing (5-8). However, the methods mentioned above have limited ability for understanding the M. pneumoniae epidemiological investigations. (14). Bacterial DNA from cultivated M. pneumoniae isolates was extracted using the QIAamp DNA minikit (Qiagen), and the aliquots were placed in a refrigerator at Ϫ20°C. p1 gene-based genotyping of M. pneumoniae. Sixty of 201 M. pneumoniae clinical isolates were genotyped by full-length sequencing of the p1 gene in our previous report (8); the other 141 isolates were genotyped by a previously reported method (15, 16). Results indicated that 184 (91.5%) isolates were type 1, 2 (1.0%) isolates were variant 2a, 15 (7.5%) isolates were variant 2c, and no type 2 isolates were detected.Sequencing analysis of the 23S rRNA gene. The domain V of 23S rRNA, related to M. pneumoniae macrolide resistance, was detected using a previously reported method (17). Results indicated that 24 (11.9%) isolates were macrolide sensitive without mutation and 177 (88.1%) isolates were macrolide resistant with mutation within the domain V of 23S rRNA, of which the numbers of A2063G, A2064G, and A2064T mutant isolates were 171, 5, and 1, respectively. M. pneumoniae MLVA genotyping. A total of 201 M. pneumoniae clinical isolates were analyzed using the MLVA assay (9), with M129 as a reference strain. Using nonfluorescent primers, PCR amplification against 5 VNTRs from 10 randomly selected isolates was performed to validate the creditability of MLVA genotyping. All data were analyzed using BioNumerics (version 6.5) software. Results indicated that 201 M. pneumoniae clinical isolates were divided into 16 MLVA types, of which 10 types were reported previously, and the other 6 types were new MLVA typ...

show abstract

Genotyping analysis of Mycoplasma pneumoniae clinical strains in Japan between 1995 and 2005: type shift phenomenon of M. pneumoniae clinical strains

Cited by 87 publications

References 25 publications

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

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

Comparison of Commercial and In-House Real-Time PCR Assays Used for Detection of Mycoplasma pneumoniae

Multiple-Locus Variable-Number Tandem-Repeat Analysis of 201 Mycoplasma pneumoniae Isolates from Beijing, China, from 2008 to 2011

Contact Info

Product

Resources

About