MOLECULAR EPIDEMIOLOGY, VIRULENCE, AND GENOMIC CHARACTERIZATION OF UREAPLASMAS
Ken B Waites
University Of Alabama At Birmingham, 1530 3rd Avenue South, Birmingham, Al 35294
Grant 5R01AI072577-04 from National Institute Of Allergy And Infectious Diseases
Abstract: Ureaplasma spp. colonize many healthy persons, yet they may also cause invasive diseases. Reasons they are commensals in some instances and produce systemic infections in others are unknown. There is increasing evidence that some Ureaplasma serovars may have a greater pathogenic potential than others. However, proof of this concept is incomplete. Prior attempts to study pathogenesis were hampered by imprecise typing methods, cross-reactions, lack of commercial reagents, and the fact that multiple serovars may be present simultaneously. Contradictory findings regarding differential pathogenicity of the 2 Ureaplasma species and individual serovars also suggests the possibility there may be virulence factors that were not detected using older, less discriminatory techniques. We hypothesize that differential pathogenicity of Ureaplasma spp. may be explained by analyzing clinical isolates and 14 serovars genotypically to identify genetic differences and possibly dissimilar expression of virulence factors. This research will examine Ureaplasma spp. from persons with invasive infections and compare them with others from persons without these conditions. Our Specific Aims are to (1) Determine occurrence of Ureaplasma species and serovars in clinical isolates from a variety of different conditions and in commensal organisms using PCR; (2) Refine and further develop pulsed field gel electrophoresis (PFGE) and restriction fragment length polymorphism for use in determining genetic relatedness between Ureaplasma serovars and within serovars of pathogenic versus commensal isolates; (3) Compare the size and number of tandem amino acid repeats in multiple banded antigens of pathogenic versus commensal isolates a means to assess this characteristic as a predictor of disease; (4) Identify genes that encode potential virulence factors lgA1 protease and phospholipase A1, A2, and C activities using global transposon mutagenesis and comparative genomic analysis of all 14 serovars and selected clinical isolates to search for likely candidates for these genes, followed by cloning and expressing the genes to determine if their presence or expression correlates with pathogenic outcome. Identification of distinctive features of invasive strains may guide development of diagnostic tools and guide future studies to improve understanding of diseases affecting vulnerable populations including pregnant women and their infants, preventive strategies, and management
Keywords: 21+ years old; ATGN; Achievement; Achievement Attainment; Address; Adult; Affect; Amino Acids; Antigens; Arthritis; BPD; Bacteremia; Bronchopulmonary Dysplasia; Candidate Disease Gene; Candidate Gene; Characteristics; Chorioamnionitis; Clinical; Clinical Research; Clinical Study; Cloning; Collection; Comparative Genomic Analysis; Cross Reactions; Development; Diagnostic; Disease; Disorder; Electrophoresis, Gel, Pulsed-Field; Electrophoresis, Gel, Pulsed-Field Gradient; Endometritis; Environmental Factor; Environmental Risk Factor; Epidemiology, Molecular; Esteroproteases; Fractionation, Pulse Field; Future; Gene variant; Genes; Genetic; Genetic Diversity; Genetic Variation; Genetics-Mutagenesis; Genitourinary; Genitourinary system; Genomics; Goals; Human; Human, Adult; Human, General; IgA; Immune system; Immunoglobulin A; Individual; Infant; Infant, Premature; Infection; Infection of amniotic sac and membranes; Investigators; Knowledge; Lecithinase A1; Lecithinases; Low Birth Weight Infant; Lower respiratory tract structure; Man (Taxonomy); Man, Modern; Meningitis; Methods; Methods and Techniques; Methods, Other; Microbe Genome Sequencing; Microbial Genome Sequencing; Miscarriage; Molecular Biology, Mutagenesis; Molecular Biology, Restriction Fragment Length Polymorphisms; Molecular Epidemiology; Mutagenesis; NIAID; National Institute of Allergy and Infectious Disease; Organism; Outcome; PFGE; Pathogenesis; Pathogenicity; Pathogenicity Factors; Peptidases; Peptide Hydrolases; Persons; Phosphatidate 1-Acylhydrolase; Phospholipase; Phospholipase A1; Pneumonia; Pneumonitis; Predisposition; Pregnancy Outcome; Pregnant Women; Premature Infant; Prevention; Prevention strategy; Preventive strategy; Programs (PT); Programs [Publication Type]; Proteases; Proteinases; Proteolytic Enzymes; Pulmonary Inflammation; RFLP; Reagent; Research; Research Personnel; Researchers; Restriction Fragment Length Polymorphism Analysis; Restriction fragment length polymorphism; Spontaneous abortion; Susceptibility; Systemic disease; Systemic infection; T-Mycoplasma; Techniques; Tetracycline Resistance; Tetracycline Resistant; Time; Ureaplasma; Ureaplasma Infections; Urethritis; Urinary Calculi; Urinary Stones; Urinary Tract Stones; Urogenital; Urogenital System; Urolith; VLBW (human); Variation (Genetics); Virulence; Virulence Factors; Vulnerable Populations; Woman; Work; adult human (21+); allelic variant; aminoacid; arthritic; bacteraemia; base; body system, allergic/immunologic; chronic lung disease in infants; chronic lung disease in neonatal infants; chronic lung disease in neonates; chronic lung disease in newborns; chronic lung disease in prematurity; design; designing; disease/disorder; environmental risk; experiment; experimental research; experimental study; immunogen; improved; infant chronic lung disease; innovate; innovation; innovative; living system; low birth weight infant human; low birthweight; lower respiratory tract; men; men`s; mutant; neonatal chronic lung disease; neonate; newborn chronic lung disease; organ system, allergic/immunologic; premature; premature baby; premature infant human; preterm baby; preterm infant; preterm infant human; preterm neonate; programs; research study; sex; tool; ureaplasma infected; urogenital system (urinary part)
Project start date: 2007-03-15
Project end date: 2011-02-28
Budget start date: 1-MAR-2010
Budget end date: 28-FEB-2011
5R01AI072577-04 (2010): $427332
Sponsored Links Excellgen http://Excellgen.com
Molecular Epidemiology, Virulence, And Genomic Characterization Of Ureaplasmas
Ken B Waites, Medical Director
Pathologyuniversity Of Alabama At Birmingham
Grant 5R01AI072577-03 from National Institute Of Allergy And Infectious Diseases IRG: CRFS
Abstract: Ureaplasma spp. colonize many healthy persons, yet they may also cause invasive diseases. Reasons they are commensals in some instances and produce systemic infections in others are unknown. There is increasing evidence that some Ureaplasma serovars may have a greater pathogenic potential than others. However, proof of this concept is incomplete. Prior attempts to study pathogenesis were hampered by imprecise typing methods, cross-reactions, lack of commercial reagents, and the fact that multiple serovars may be present simultaneously. Contradictory findings regarding differential pathogenicity of the 2 Ureaplasma species and individual serovars also suggests the possibility there may be virulence factors that were not detected using older, less discriminatory techniques. We hypothesize that differential pathogenicity of Ureaplasma spp. may be explained by analyzing clinical isolates and 14 serovars genotypically to identify genetic differences and possibly dissimilar expression of virulence factors. This research will examine Ureaplasma spp. from persons with invasive infections and compare them with others from persons without these conditions. Our Specific Aims are to (1) Determine occurrence of Ureaplasma species and serovars in clinical isolates from a variety of different conditions and in commensal organisms using PCR; (2) Refine and further develop pulsed field gel electrophoresis (PFGE) and restriction fragment length polymorphism for use in determining genetic relatedness between Ureaplasma serovars and within serovars of pathogenic versus commensal isolates; (3) Compare the size and number of tandem amino acid repeats in multiple banded antigens of pathogenic versus commensal isolates a means to assess this characteristic as a predictor of disease; (4) Identify genes that encode potential virulence factors lgA1 protease and phospholipase A1, A2, and C activities using global transposon mutagenesis and comparative genomic analysis of all 14 serovars and selected clinical isolates to search for likely candidates for these genes, followed by cloning and expressing the genes to determine if their presence or expression correlates with pathogenic outcome. Identification of distinctive features of invasive strains may guide development of diagnostic tools and guide future studies to improve understanding of diseases affecting vulnerable populations including pregnant women and their infants, preventive strategies, and management
Keywords: Mycoplasmatales, epidemiology, virulence acid, antigen, arthritis, base, bronchopulmonary dysplasia, bronchus, concept, conditioning, disease /disorder prevention /control, emotion, endopeptidase, gene, genetics, genome, human, hypogammaglobulinemia, immune system, infection, lung, meningitis, molecular cloning, mutant, organism, pneumonia, pregnancy, pulsed field gel electrophoresis, restriction fragment length polymorphism, septicemia, sex, spontaneous abortion, tetracycline, urethritis, urinary calculi clinical research
Project start date: 2007-03-15
Project end date: 2011-02-28
Grants awarded to Ken B Waites
Molecular Epidemiology, Virulence, And Genomic Characterization Of Ureaplasmas
Ken B Waites, Medical Director
Pathologyuniversity Of Alabama At Birmingham, 1530 3rd Avenue South, Birmingham, Al 35294
Grant 1R01AI072577-01 from National Institute Of Allergy And Infectious Diseases IRG: CRFS
Abstract: DESCRIPTION () Ureaplasma spp. colonize many healthy persons, yet they may also cause invasive diseases. Reasons they are commensals in some instances and produce systemic infections in others are unknown. There is increasing evidence that some Ureaplasma serovars may have a greater pathogenic potential than others. However, proof of this concept is incomplete. Prior attempts to study pathogenesis were hampered by imprecise typing methods, cross-reactions, lack of commercial reagents, and the fact that multiple serovars may be present simultaneously. Contradictory findings regarding differential pathogenicity of the 2 Ureaplasma species and individual serovars also suggests the possibility there may be virulence factors that were not detected using older, less discriminatory techniques. We hypothesize that differential pathogenicity of Ureaplasma spp. may be explained by analyzing clinical isolates and 14 serovars genotypically to identify genetic differences and possibly dissimilar expression of virulence factors. This research will examine Ureaplasma spp. from persons with invasive infections and compare them with others from persons without these conditions. Our Specific Aims are to (1) Determine occurrence of Ureaplasma species and serovars in clinical isolates from a variety of different conditions and in commensal organisms using PCR; (2) Refine and further develop pulsed field gel electrophoresis (PFGE) and restriction fragment length polymorphism for use in determining genetic relatedness between Ureaplasma serovars and within serovars of pathogenic versus commensal isolates; (3) Compare the size and number of tandem amino acid repeats in multiple banded antigens of pathogenic versus commensal isolates a means to assess this characteristic as a predictor of disease; (4) Identify genes that encode potential virulence factors lgA1 protease and phospholipase A1, A2, and C activities using global transposon mutagenesis and comparative genomic analysis of all 14 serovars and selected clinical isolates to search for likely candidates for these genes, followed by cloning and expressing the genes to determine if their presence or expression correlates with pathogenic outcome. Identification of distinctive features of invasive strains may guide development of diagnostic tools and guide future studies to improve understanding of diseases affecting vulnerable populations including pregnant women and their infants, preventive strategies, and management.
Project start date: 2007-03-15
Project end date: 2011-02-28
1R01AI072577-01 (2007): $460917