-
Molecular Microbiology Feb 2004Mycoplasma pneumoniae is a minimal microbe with respect to cell envelope composition, biosynthetic and regulatory capabilities and genome size, yet it possesses a... (Review)
Review
Mycoplasma pneumoniae is a minimal microbe with respect to cell envelope composition, biosynthetic and regulatory capabilities and genome size, yet it possesses a remarkably complex, multifunctional terminal organelle. This membrane-bound extension of the mycoplasma cell is defined by the presence of an electron-dense core that appears as paired, parallel bars oriented longitudinally and enlarging at the distal end to form a terminal button. Most non-cytadhering mutants of M. pneumoniae isolated to date exhibit defects in the architecture of the terminal organelle. Detailed characterization of those mutants has revealed the identities of many component proteins of the terminal organelle as well as the likely order in which some of those components are required. Additional questions regarding the composition of the electron-dense core, the means by which the terminal organelle is duplicated during cell division and the manner in which this process is regulated remain to be answered. Thus, it seems that there is much to be learned about cellular engineering and spatial regulation in these 'simple' cell wall-less bacteria.
Topics: Adhesins, Bacterial; Bacterial Adhesion; Genes, Reporter; Green Fluorescent Proteins; Humans; Luminescent Proteins; Mycoplasma pneumoniae; Organelles; Pneumonia, Mycoplasma; Respiratory Mucosa
PubMed: 14763969
DOI: 10.1046/j.1365-2958.2003.03899.x -
Clinical Microbiology and Infection :... Mar 2019Rapid detection of macrolide resistance-associated mutations in Mycoplasma pneumoniae is crucial for effective antimicrobial treatment. We evaluated the Lightmix...
OBJECTIVES
Rapid detection of macrolide resistance-associated mutations in Mycoplasma pneumoniae is crucial for effective antimicrobial treatment. We evaluated the Lightmix Mycoplasma macrolide assay for the detection of point mutations at nucleotide positions 2063 and 2064 in the 23S ribosomal RNA (rRNA) gene of M. pneumoniae that confer macrolide resistance.
METHODS
Samples from 3438 patients with a respiratory tract infection were analysed by M. pneumoniae real-time PCR, and 208 (6%) of them were tested positive. In this retrospective study, 163 M. pneumoniae real-time PCR-positive samples were analysed by the Lightmix assay, and results were compared to targeted 23S rRNA sequencing.
RESULTS
Macrolide-resistant M. pneumoniae were found in 15 (9%) of 163 retrospectively analysed samples. The Lightmix assay showed a sensitivity of 100% (95% confidence interval, 78.2-100) and a specificity of 100% (95% confidence interval, 97.5-100) as the detected M. pneumoniae genotype (148 wild type and 15 non-wild type) was confirmed by 23S rRNA sequencing in all samples.
CONCLUSIONS
The Lightmix assay is an easy-to-use and accurate molecular test that allows rapid determination of macrolide resistance in M. pneumoniae.
Topics: Anti-Bacterial Agents; DNA, Bacterial; Drug Resistance, Bacterial; Genotype; Humans; Macrolides; Mycoplasma pneumoniae; Pneumonia, Mycoplasma; Point Mutation; RNA, Ribosomal, 23S; Real-Time Polymerase Chain Reaction; Retrospective Studies; Sensitivity and Specificity; Sequence Analysis, DNA
PubMed: 30391582
DOI: 10.1016/j.cmi.2018.10.006 -
BMC Genomics Aug 2015Mycoplasma pneumoniae is a common pathogen that causes upper and lower respiratory tract infections in people of all ages, responsible for up to 40% of...
BACKGROUND
Mycoplasma pneumoniae is a common pathogen that causes upper and lower respiratory tract infections in people of all ages, responsible for up to 40% of community-acquired pneumonias. It also causes a wide array of extrapulmonary infections and autoimmune phenomena. Phylogenetic studies of the organism have been generally restricted to specific genes or regions of the genome, because whole genome sequencing has been completed for only 4 strains. To better understand the physiology and pathogenicity of this important human pathogen, we performed comparative genomic analysis of 15 strains of M. pneumoniae that were isolated between the 1940s to 2009 from respiratory specimens and cerebrospinal fluid originating from the USA, China and England.
RESULTS
Illumina MiSeq whole genome sequencing was performed on the 15 strains and all genome sequences were completed. Results from the comparative genomic analysis indicate that although about 1500 SNP and indel variants exist between type1 and type 2 strains, there is an overall high degree of sequence similarity among the strains (>99% identical to each other). Within the two subtypes, conservation of most genes, including the CARDS toxin gene and arginine deiminase genes, was observed. The major variation occurs in the P1 and ORF6 genes associated with the adhesin complex. Multiple hsdS genes (encodes S subunit of type I restriction enzyme) with variable tandem repeat copy numbers were found in all 15 genomes.
CONCLUSIONS
These data indicate that despite conclusions drawn from 16S rRNA sequences suggesting rapid evolution, the M. pneumoniae genome is extraordinarily stable over time and geographic distance across the globe with a striking lack of evidence of horizontal gene transfer.
Topics: China; Comparative Genomic Hybridization; England; Evolution, Molecular; Genetic Variation; Genome, Bacterial; High-Throughput Nucleotide Sequencing; Humans; Mycoplasma pneumoniae; Phylogeny; Sequence Analysis, DNA; Sequence Homology, Nucleic Acid; United States
PubMed: 26275904
DOI: 10.1186/s12864-015-1801-0 -
International Journal of Medical... Apr 2018Mycoplasma pneumoniae is a bacterial pathogen of humans that is a major causative agent of chronic respiratory disease. M. pneumoniae infections often recur even after...
Mycoplasma pneumoniae is a bacterial pathogen of humans that is a major causative agent of chronic respiratory disease. M. pneumoniae infections often recur even after successful treatment of symptoms with antibiotics, and resistance to antibiotics is increasing worldwide, with nearly complete resistance in some places. Although biofilms often contribute to chronicity and resistance, M. pneumoniae biofilms remain poorly characterized. Scanning electron microscopy revealed that cells of wild-type (WT) M. pneumoniae strain M129 biofilms, as well as mutants II-3 and II-3R, in vitro became increasingly rounded as the biofilm towers matured over 5 days. The role of gliding motility in biofilm formation was addressed by analyzing differences in biofilm architecture in non-motile mutant II-3R and hypermotile mutant prpC-and by using time-lapse microcinematography to measure flux of cells around biofilm towers. There were no major differences in biofilm architecture between WT and motility mutants, with perhaps a slight tendency for the prpC- cells to spread outside towers during early stages of biofilm formation. Consistent with an insignificant role of motility in biofilm development, flux of cells near towers, which was low, was dominated by exit of cells. Immunofluorescence microscopy revealed that motility-associated attachment organelle (AO) proteins exhibited no discernable changes in localization to foci over time, but immunoblotting identified a decrease in steady-state levels of protein P200, which is required for normal gliding speed, as the WT culture aged. Non-adherent strain II-3 and non-motile strain II-3R also exhibited a steady decrease in P200 steady-state levels, suggesting that the decrease in P200 levels was not a response to changes in gliding behavior during maturation. We conclude that M. pneumoniae cells undergo morphological changes as biofilms mature, motility plays no major role in biofilm development, and P200 loss might be related to maturation of cells. This study helps to characterize potential therapeutic targets for M. pneumoniae infections.
Topics: Bacterial Adhesion; Biofilms; Humans; In Vitro Techniques; Microscopy, Electron, Scanning; Mycoplasma pneumoniae
PubMed: 29426802
DOI: 10.1016/j.ijmm.2018.01.007 -
Methods of Biochemical Analysis 2006
Review
Topics: Bacterial Proteins; Detergents; Electrophoresis, Gel, Two-Dimensional; Genes, Bacterial; Mycoplasma pneumoniae; Octoxynol; Open Reading Frames; Phosphoproteins; Protein Processing, Post-Translational; Protein Sorting Signals; Proteome; Proteomics; RNA
PubMed: 16929672
DOI: No ID Found -
International Journal of Infectious... Jun 2019Analysis of the molecular characteristics of isolates is very important for clinical and epidemiological study of community-acquired pneumonia (CAP) caused by Mycoplasma...
BACKGROUND
Analysis of the molecular characteristics of isolates is very important for clinical and epidemiological study of community-acquired pneumonia (CAP) caused by Mycoplasma pneumoniae.
METHODS
Between 2010 and 2012, an epidemic period, M. pneumoniae was isolated from oropharyngeal swabs of consecutive CAP patients. Minimum inhibitory concentrations of macrolides, 23S rRNA gene sequencing, P1 gene and multilocus variable-number tandem-repeat analysis (MLVA) genotyping was conducted.
RESULTS
88.3% (181/205) of the isolates were macrolide-resistant M. pneumoniae (MRMP) and all harbored an A2063 G mutation. The strains were clustered into 7 MLVA types, and P1 type 1 and type 2 lineages were co-circulated (86.3% and 13.7%). Compared with adults, no specific MLVA type contributed to higher M. pneumoniae infection in children (p = 0.14). Similar macrolide profile and genotypes of M. pneumoniae was found between outpatients and inpatients. Significant differences in proportion of P1 types and two main MLVA types 4/5/7/2 and 3/5/6/2 were observed between MRMP and macrolide-sensitive M. pneumoniae (MSMP) (p < 0.001).
CONCLUSIONS
This study demonstrates a comprehensive profile of M. pneumoniae molecular characterization among CAP patients of all age, and provides more evidences on a correlation between MLVA type 4/5/7/2 and macrolide resistance in the setting of high incidence of MRMP.
Topics: Adult; Aged; Child; Child, Preschool; China; Community-Acquired Infections; Drug Resistance, Bacterial; Epidemics; Female; Humans; Infant; Macrolides; Male; Middle Aged; Mycoplasma pneumoniae; Pneumonia, Mycoplasma; Young Adult
PubMed: 30926541
DOI: 10.1016/j.ijid.2019.03.028 -
Acta Microbiologica Et Immunologica... Mar 2013Mycoplasma pneumoniae (M. pneumoniae) is one of the most important pathogens that cause respiratory tract infection in children and adults. In this study, we describe a...
Mycoplasma pneumoniae (M. pneumoniae) is one of the most important pathogens that cause respiratory tract infection in children and adults. In this study, we describe a rapid and sensitive colorimetric loop mediated isothermal amplification (LAMP) method to detect M. pneumoniae. The specificity and sensitivity of this assay were detected with 21 common respiratory pathogens and 39 M. pneumoniae DNA. The sensitivity of LAMP was 100% among 39 M. pneumoniae isolates and the specificity was 100% among 9 members of other Mycoplasma and 12 common respiratory pathogens. The lowest detectable limit (LDL) of this assay was 102 copies, which detected by a series of standard M. pneumoniae DNA. To evaluate the clinical applicability of the LAMP assay, a total of 80 clinical samples were examined by conventional PCR, real-time PCR and the LAMP assays, respectively. The positive rates were 15.0%, 32.5% and 26.3%, respectively. This colorimetric LAMP assay demonstrated a high level of sensitivity comparable with that of conventional PCR for the detection of M. pneumoniae. It is a valuable method for simple, cost-effective and rapid detection of M. pneumoniae in the rural areas and basic clinical of China.
Topics: Colorimetry; Humans; Mycoplasma pneumoniae; Nucleic Acid Amplification Techniques; Sensitivity and Specificity
PubMed: 23529294
DOI: 10.1556/AMicr.60.2013.1.1 -
CMAJ : Canadian Medical Association... Oct 2019
Topics: Adult; Amoxicillin-Potassium Clavulanate Combination; Azithromycin; Cough; Erythema Multiforme; Fatigue; Female; Humans; Mycoplasma pneumoniae; Prednisone
PubMed: 31659061
DOI: 10.1503/cmaj.190850 -
Microbiology (Reading, England) Oct 2011The attachment organelles of bacterial species belonging to the Mycoplasma pneumoniae phylogenetic cluster are required for host cytadherence, gliding motility and...
The attachment organelles of bacterial species belonging to the Mycoplasma pneumoniae phylogenetic cluster are required for host cytadherence, gliding motility and virulence. Despite being closely related, these bacteria possess distinct cellular morphologies and gliding characteristics. The molecular mechanisms for most attachment organelle phenotypes, including shape and ability to power motility, are obscure. The attachment organelle-associated P30 protein of M. pneumoniae is implicated in both adherence and motility, with mutations negatively impacting cell morphology, adherence, gliding and virulence. To test whether the P30 alleles of different mycoplasma species confer species-specific attachment organelle properties, we created an M. pneumoniae strain in which the Mycoplasma genitalium P30 orthologue, P32, was substituted for the native P30. Selected clones were visualized by scanning electron microscopy to assess morphology and by indirect immunofluorescence microscopy to localize P32. Cytadherence ability and gliding motility were assessed by haemadsorption assay and phase-contrast microcinematography, respectively. Cell and attachment organelle morphologies were indistinguishable from wild-type M. pneumoniae as well as M. pneumoniae II-3 expressing a C-terminally 6×His-tagged P30 construct. P32 was localized to the tip of the attachment organelle of transformant cells. Although a specific role for P30 in species-specific phenotypes was not identified, this first test of orthologous gene replacement in different mycoplasma species demonstrates that the differences in the M. pneumoniae and M. genitalium proteins contribute little if anything to the different attachment organelle phenotypes between these species.
Topics: Adhesins, Bacterial; Amino Acid Sequence; Humans; Molecular Sequence Data; Mycoplasma genitalium; Mycoplasma pneumoniae; Organelles; Protein Transport; Sequence Alignment
PubMed: 21778204
DOI: 10.1099/mic.0.052464-0 -
European Journal of Clinical... Aug 2021Mycoplasma pneumoniae (M. pneumoniae) is an important pathogen in community-acquired pneumonia. The community-acquired respiratory distress syndrome (CARDS) toxin is the...
Mycoplasma pneumoniae (M. pneumoniae) is an important pathogen in community-acquired pneumonia. The community-acquired respiratory distress syndrome (CARDS) toxin is the only known virulence factor of M. pneumoniae. It is worth exploring whether this toxin can be used as a candidate antigen for the serodiagnosis of M. pneumoniae. In this study, the full-length, N-terminal, and C-terminal regions of the CARDS toxin were expressed and purified, and serological reactions were evaluated using ELISA. A total of 184 serum samples were collected and tested using a commercialized test kit. Eighty-seven samples were positive, and 97 samples were negative for infection. The purified recombinant proteins were used as antigens to test the serum via indirect ELISA. The sensitivity of the CARDS toxin, the N-terminal region, and the C-terminal region were 90.8%, 90.8%, and 92.0%, respectively. The specificity of the CARDS toxin, the N-terminal region, and the C-terminal region were 85.6%, 73.2%, and 93.8%, respectively. All three CARDS toxin proteins exhibited good reactivity, of which the C-terminal region had a good discrimination ability in human sera. This may have a potential diagnostic value for M. pneumoniae infections.
Topics: Bacterial Proteins; Bacterial Toxins; Enzyme-Linked Immunosorbent Assay; Gene Expression Regulation, Bacterial; Humans; Mycoplasma pneumoniae; Pneumonia, Mycoplasma; Sensitivity and Specificity; Serologic Tests
PubMed: 33733396
DOI: 10.1007/s10096-021-04209-2