DOCSLIB.ORG

Microbiology Legend Cycle 29 Organims 6 Mycoplasma

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Microbiology Legend Cycle 29 Organims 6 Mycoplasma P.O. Box 131375, Bryanston, 2074 Ground Floor, Block 5 Bryanston Gate, Main Road Bryanston, Johannesburg, South Africa www.thistle.co.za Tel: +27 (011) 463 3260 Fax: +27 (011) 463 3036 Fax to Email: + 27 (0) 86‐538‐4484 e‐mail : [email protected] Please read this section first The HPCSA and the Med Tech Society have confirmed that this clinical case study, plus your routine review of your EQA reports from Thistle QA, should be documented as a “Journal Club” activity. This means that you must record those attending for CEU purposes. Thistle will not issue a certificate to cover these activities, nor send out “correct” answers to the CEU questions at the end of this case study. The Thistle QA CEU No is: MT00025. Each attendee should claim THREE CEU points for completing this Quality Control Journal Club exercise, and retain a copy of the relevant Thistle QA Participation Certificate as proof of registration on a Thistle QA EQA. MICROBIOLOGY LEGEND CYCLE 29 ORGANIMS 6 MYCOPLASMA Mycoplasma, the smallest organisms capable of self-replication was first isolated by Roux and Nocard from cattle with contagious bovine pleuropneumonia in 1898. Subsequent isolates from other sources were termed pleuro-pneumonia-like organisms. Members of the class Mollicutes (Latin: mollis means soft and cutin skin), they are characterised by their absence of a cell wall and are therefore refractory to treatment with β- lactam class antibiotics. All Mycoplasma require lipid precursors for plasma membrane synthesis. Mycoplasma is a species of gram-negative bacteria that is a common inhabitant of the human oropharynx and also a cell culture contaminant. It is usually non-pathogenic but it can cause infections in leukemic or immunosuppressed patients. Like viruses they pass through filters that retain bacteria, but except for the cell wall, possess all other cellular constituents of bacteria. Unlike cell-wall deficient bacteria, they fail to revert to bacteria when cultured under appropriate conditions. Despite superficial cultural similarities, the Mycoplasma comprise a heterogenous group of micro-organisms differing from each other in DNA composition, metabolic reactions, nutritional requirements, antigenic composition and host specie specificity. They are divided into 3 taxonomic families: The Mycoplasmataceae and Spiroplasmataceae, both requiring cholesterol for growth and non-cholesterol-dependent Acholeplasmataceae. Included in the family Mycoplasmataceae are the Mycoplasma species which do not hydrolyse urea and the genus Ureaplasma, strains of which produce abundant quantities of urease. There are 51 recognised species in the genus Mycoplasma. 10 Mycoplasma species have been detected in the clinical laboratory. Mycoplasma pneumonia is the cause of primary atypical pneumonia. Mycoplasma salivarian and Mycoplasma orale are common inhabitants of the oropharynx, particularly in individuals with poor oral hygiene, whereas Mycoplasma buccale is a less frequent oropharyngeal colonizer. Mycoplasma is ubiquitous in nature as saprophytes and/or parasites of plants and animals. Mycoplasma species also cause contamination of cell cultures, interfering with their metabolic processes and slowing their rate of growth. Thistle QA is a SANAS accredited organisation, No: PTS0001 Accredited to ISO guide 43 and ILAC G13 Certificate available on request or at www.sanas.co.za P.O. Box 131375, Bryanston, 2074 Ground Floor, Block 5 Bryanston Gate, Main Road Bryanston, Johannesburg, South Africa www.thistle.co.za Tel: +27 (011) 463 3260 Fax: +27 (011) 463 3036 Fax to Email: + 27 (0) 86‐538‐4484 e‐mail : [email protected] Mycoplasma infection in a cell culture Mycoplasma orale incubation for seven days on Mycoplasma Experience solid Mycoplasma medium The Mycoplasma appears to be host-specific, being communicable and potentially pathogenic only within a single host species. Mycoplasma has been cultivated from human mucous membranes and tissues, particularly from the genital, urinary and respiratory tracts. Mycoplasma is part of the normal flora of the mouth and can be grown from normal saliva, oral mucous membranes, sputum or tonsillar tissue. M. salivarium, M. orale and other Mycoplasma can be recovered from the oral cavities of many healthy adults, but an association with clinical disease is uncertain. M. hominis is found in the oropharynx of less than 5% of adults. M pneumonia in the oropharynx is generally associated with disease. Some Mycoplasma are inhabitants of the genitourinary tract, particularly in females. In both men and women, genital carriage of Mycoplasma is directly related to the number of lifetime sex partners. Diagnostic Laboratory Tests Specimens – consists of throat swabs, sputum, inflammatory exudates and respiratory, urethral or genital secretions. Microscopic examination – direct examination of a specimen for Mycoplasma is useless. Cultures – the material is inoculated onto special solid media and incubated for 3-10 days at 37°C with 5% CO2 (under micro-aerophilic conditions), or in special broth and incubated aerobically. One or two transfers of media may be necessary before growth appears that is suitable for microscopic examination by staining or immuno-fluorescence. Colonies may have a “fried egg” appearance on agar. Serology – antibodies develop in humans infected with Mycoplasma and can be demonstrated by several methods. Treatment Many strains of Mycoplasma are inhibited by a variety of antimicrobial drugs, but most strains are resistant to penicillin, cephalosporin and vancomycin. Tetracycline and erythromycin are effective both in vitro and in vivo and are, at present, the drugs of choice in mycoplasmal pneumonia. References 1. http://en.wikipedia.org/wiki/Mycoplasma 2. Clinical laboratory medicine, Volume 2001, Kenneth D. McClatchey 3. Medical Microbiology 20th edition, Brooks, Butel & Ornston Questions 1. Discuss the morphological characteristics of the Mycoplasma. 2. Discuss the pathophysiology of Mycoplasma. 3. Discuss the diagnostic laboratory tests that can be used to identify this organism. Thistle QA is a SANAS accredited organisation, No: PTS0001 Accredited to ISO guide 43 and ILAC G13 Certificate available on request or at www.sanas.co.za .
Load more

Recommended publications
  • Mycoplasma Agalactiae MEMBRANE PROTEOME
    UNIVERSITÀ DEGLI STUDI DI SASSARI SCUOLA DI DOTTORATO IN SCIENZE BIOMOLECOLARI E BIOTECNOLOGICHE INDIRIZZO MICROBIOLOGIA MOLECOLARE E CLINICA XXIII Ciclo CHARACTERIZATION OF Mycoplasma agalactiae MEMBRANE PROTEOME Direttore: Prof. Bruno Masala Tutor: Dr. Alberto Alberti Tesi di dottorato della Dott.ssa Carla Cacciotto ANNO ACCADEMICO 2009-2010 TABLE OF CONTENTS 1. Abstract 2. Introduction 2.1 Mycoplasmas: taxonomy and main biological features 2.2 Metabolism 2.3 In vitro cultivation 2.4 Mycoplasma lipoproteins 2.5 Invasivity and pathogenicity 2.6 Diagnosis of mycoplasmosis 2.7 Mycoplasma agalactiae and Contagious Agalactia 3. Research objectives 4. Materials and methods 4.1 Media and buffers 4.2 Bacterial strains and culture conditions 4.3 Total DNA extraction and PCR 4.4 Total proteins extraction 4.5 Triton X-114 fractionation 4.6 SDS-PAGE 4.7 Western immunoblotting 4.8 2-D PAGE 4.9 2D DIGE 4.10 Spot picking and in situ tryptic digestion 4.11 GeLC-MS/MS 4.12 MALDI-MS 4.13 LC-MS/MS 4.14 Data analysis Dott.ssa Carla Cacciotto, Characterization of Mycoplasma agalactiae membrane proteome. Tesi di Dottorato in Scienze Biomolecolari e Biotecnologiche, Università degli Studi di Sassari. 5. Results 5.1 Species identification 5.2 Extraction of bacterial proteins and isolation of liposoluble proteins 5.3 2-D PAGE/MS of M. agalactiae PG2T liposoluble proteins 5.4 2D DIGE of liposoluble proteins among the type strain and two field isolates of M. agalactiae 5.5 GeLC-MS/MS of M. agalactiae PG2T liposoluble proteins 5.6 Data analysis and classification 6. Discussion 7.
    [Show full text]
  • (12) United States Patent (10) Patent No.: US 8,835,117 B2 Mitchell Et Al
    US008835117B2 (12) United States Patent (10) Patent No.: US 8,835,117 B2 Mitchell et al. (45) Date of Patent: Sep. 16, 2014 (54) NUCLEICACIDS FOR DETECTION AND OTHER PUBLICATIONS DISCRIMINATION OF GENOTYPES OF CHLAMYDOPHILA PSITTAC Nazarenko, I. Methods in Molecular Biology (2006) 335: 95-114.* Jeffrey et al. Microbiology (2007) 153: 2679-2688.* 75) Inventors: Stephaniep L. Mitchell, Somerville, MA Geens et al. Journal of Clinical Microbiology (2005) 43(5): 2456 (US); Jonas M. Winchell, Lilburn, GA 2461. (US) Geens et al., “Development of a Chlamydophila psittaci species specific and genotype-specific real-time PCR.” Vet. Res., 36: 787 (73) Assignee: The United States of America as 797, 2005. represented by the Secretary of the Geens et al., “Sequencing of the Chlamydophila psittaci OmpA Gene Department of Health and Human Reveals a New Genotype, E/B, and the Need for a Rapid Discrimi Services, Centers for Disease Control natory Genotyping Method.” J. Clin. Microbiol. 43(5): 2456-2461, and Prevention, Washington, DC (US) 2005. Heddema, “Genotyping of Chlamydophila psittaci in Human (*) Notice: Subject to any disclaimer, the term of this Samples.” Emerging Infectious Diseases, 12(12): 1989-1990, 2006. patent is extended or adjusted under 35 Menard, “Development of a real-time PCR for the detection of U.S.C. 154(b) by 82 days. Chlamydia psittaci,” J. Med. Microbiol. 55(Pt. 4): 471-473, 2006. Mitchellet al., “Genotyping of Chlamydophilapsittaci by Real-Time (21) Appl. No.: 13/322,787 PCR and High-Resolution Melt Analysis,” Journal of Clinical Microbiology, 47(1): 175-181, 2009. (22) PCT Filed: May 28, 2010 Sachse et al., “Genotyping of Chlamydophila psittaci using a new DNA microarray assay based on sequence analysis of ompA genes.” (86).
    [Show full text]
  • ( 12 ) United States Patent
    US009956282B2 (12 ) United States Patent ( 10 ) Patent No. : US 9 ,956 , 282 B2 Cook et al. (45 ) Date of Patent: May 1 , 2018 ( 54 ) BACTERIAL COMPOSITIONS AND (58 ) Field of Classification Search METHODS OF USE THEREOF FOR None TREATMENT OF IMMUNE SYSTEM See application file for complete search history . DISORDERS ( 56 ) References Cited (71 ) Applicant : Seres Therapeutics , Inc. , Cambridge , U . S . PATENT DOCUMENTS MA (US ) 3 ,009 , 864 A 11 / 1961 Gordon - Aldterton et al . 3 , 228 , 838 A 1 / 1966 Rinfret (72 ) Inventors : David N . Cook , Brooklyn , NY (US ) ; 3 ,608 ,030 A 11/ 1971 Grant David Arthur Berry , Brookline, MA 4 ,077 , 227 A 3 / 1978 Larson 4 ,205 , 132 A 5 / 1980 Sandine (US ) ; Geoffrey von Maltzahn , Boston , 4 ,655 , 047 A 4 / 1987 Temple MA (US ) ; Matthew R . Henn , 4 ,689 ,226 A 8 / 1987 Nurmi Somerville , MA (US ) ; Han Zhang , 4 ,839 , 281 A 6 / 1989 Gorbach et al. Oakton , VA (US ); Brian Goodman , 5 , 196 , 205 A 3 / 1993 Borody 5 , 425 , 951 A 6 / 1995 Goodrich Boston , MA (US ) 5 ,436 , 002 A 7 / 1995 Payne 5 ,443 , 826 A 8 / 1995 Borody ( 73 ) Assignee : Seres Therapeutics , Inc. , Cambridge , 5 ,599 ,795 A 2 / 1997 McCann 5 . 648 , 206 A 7 / 1997 Goodrich MA (US ) 5 , 951 , 977 A 9 / 1999 Nisbet et al. 5 , 965 , 128 A 10 / 1999 Doyle et al. ( * ) Notice : Subject to any disclaimer , the term of this 6 ,589 , 771 B1 7 /2003 Marshall patent is extended or adjusted under 35 6 , 645 , 530 B1 . 11 /2003 Borody U .
    [Show full text]
  • A Phylogenetic Analysis of the Mycoplasmas: Basis for Their Lc Assification W
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by DigitalCommons@University of Nebraska University of Nebraska - Lincoln DigitalCommons@University of Nebraska - Lincoln Public Health Resources Public Health Resources 9-1989 A Phylogenetic Analysis of the Mycoplasmas: Basis for Their lC assification W. G. Weisburg University of Illinois J. G. Tully National Institute of Allergy and Infectious Diseases D. L. Rose National Institute of Allergy and Infectious Diseases J. P. Petzel Iowa State University H. Oyaizu University of Illinois See next page for additional authors Follow this and additional works at: https://digitalcommons.unl.edu/publichealthresources Weisburg, W. G.; Tully, J. G.; Rose, D. L.; Petzel, J. P.; Oyaizu, H.; Yang, D.; Mandelco, L.; Sechrest, J.; Lawrence, T. G.; Van Etten, James L.; Maniloff, J.; and Woese, C. R., "A Phylogenetic Analysis of the Mycoplasmas: Basis for Their lC assification" (1989). Public Health Resources. 310. https://digitalcommons.unl.edu/publichealthresources/310 This Article is brought to you for free and open access by the Public Health Resources at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in Public Health Resources by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. Authors W. G. Weisburg, J. G. Tully, D. L. Rose, J. P. Petzel, H. Oyaizu, D. Yang, L. Mandelco, J. Sechrest, T. G. Lawrence, James L. Van Etten, J. Maniloff, and C. R. Woese This article is available at DigitalCommons@University of Nebraska - Lincoln: https://digitalcommons.unl.edu/ publichealthresources/310 JOURNAL OF BACTERIOLOGY, Dec. 1989, p. 6455-6467 Vol. 171, No.
    [Show full text]
  • Invivogen Insight Newsletter: Mycoplasma
    November/December 2005 An Insightful Look At InvivoGen’s Innovative Products Mycoplasma contamination remains a significant problem to Mycoplasma: The Insidious Invader of Cell Cultures the culture of mammalian cells. Mycoplasmas are the smallest and simplest and enzymatic procedures. However, none of these self-replicating organisms. Due to their seriously methods is 100% reliable. Direct growth methods are The detection of mycoplasma degraded genome they cannot perform many relatively sensitive to most species but the overall contamination is an important metabolic functions, such as cell wall production or procedure is lengthy (3 weeks), costly and less synthesis of nucleotides and amino acids. sensitive to noncultivable species. The PCR method, part of mycoplasma control and Mycoplasmas are strictly parasites. They parasitize a although rather fast and inexpensive, is limited by its should be an established wide range of organisms including humans, animals, sensitivity and the risk of positive and false negative method in every cell culture insects, and plants. results. Mycoplasma and Acholeplasma are Mollicutes, that InvivoGen has developed a new mycoplasma detection laboratory. InvivoGen is pleased comprise together more than 100 recognized species. method that promises to resolve these issues. This to introduce PlasmoTest™, a Among them, about 20 species have been described as method is based on the detection of mycoplasmas by contaminants of eukaryotic cell cultures. However engineered cells that express Toll-like receptor 2, a Mycoplasma detection kit based 5 species (Mycoplasma (M.) arginini, M. fermentans, pathogen recognition receptor that detects mycoplasmas. on a brand new technology. M. orale, M. hyorhinis and Acholeplasma laidlawii) PlasmoTest™, InvivoGen’s new mycoplasma detection are isolated in 90-95% of contaminated cell cultures1.
    [Show full text]
  • Table S1. Proportional Abundance, and Absolute Abundance of Penile Bacteria from the 266 Men Included in the HIV Seroconversion Cohort
    Table S1. Proportional abundance, and absolute abundance of penile bacteria from the 266 men included in the HIV Seroconversion Cohort. All men were uncircumcised. Proportional Abundance Absolute Abundance (16S copies/swab) Genus Median Q#1 Q#3 Median Q#1 Q#3 Prevotella 0.155 0.059 0.279 2.78E+07 9.78E+05 3.14E+08 Peptoniphilus 0.077 0.038 0.119 1.52E+07 1.66E+06 8.69E+07 Peptoniphilaceae<0.97*† 0.049 0.013 0.125 7.81E+06 2.97E+05 1.39E+08 Porphyromonas 0.032 0.010 0.067 4.47E+06 2.36E+05 9.88E+07 Finegoldia 0.029 0.005 0.104 3.37E+06 7.02E+05 2.07E+07 Corynebacterium 0.027 0.004 0.119 2.74E+06 7.12E+05 1.19E+07 Anaerococcus 0.025 0.008 0.061 4.38E+06 7.03E+05 2.10E+07 Dialister 0.018 0.002 0.045 1.76E+06 7.01E+04 3.40E+07 Lactobacillus 0.002 0.001 0.004 2.46E+05 3.33E+04 2.33E+06 Gardnerella 0.001 0.000 0.003 5.72E+04 9.27E+03 8.40E+05 Ezakiella 0.004 0.001 0.016 3.59E+05 2.59E+04 1.11E+07 Clostridiales incertae sedis XIII<0.97*† 0.003 0.000 0.010 3.80E+05 1.32E+04 1.10E+07 Mobiluncus 0.001 0.000 0.011 2.24E+05 1.18E+04 6.54E+06 Firmicutes<0.97*‡ 0.005 0.001 0.016 4.55E+05 1.76E+04 1.55E+07 Murdochiella 0.004 0.001 0.015 6.34E+05 2.70E+04 1.10E+07 Campylobacter 0.004 0.000 0.013 4.19E+05 1.69E+04 1.25E+07 Negativicoccus 0.003 0.000 0.018 3.83E+05 2.54E+04 6.11E+06 Saccharofermentans<0.97* 0.001 0.000 0.009 9.49E+04 7.09E+03 5.02E+06 Peptostreptococcus 0.004 0.000 0.027 2.13E+05 1.69E+04 1.38E+07 Proteobacteria<0.97*‡ 0.001 0.000 0.010 9.69E+04 7.26E+03 3.90E+06 Porphyromonas<0.97* 0.001 0.000 0.005 1.08E+05 4.03E+03 3.46E+06 Staphylococcus
    [Show full text]
  • Microbiology Catalogue Contents
    Microbiology catalogue Contents 3 About ATCC 4 About the ATCC / LGC Standards partnership 5 Quality control strains 7 Pharmaceutical applications 12 Food microbiology 20 Food microbiology, Genomic DNA 21 Water microbiology 24 Catalogue listing 38 Nucleic Acids prepared from ATCC Genuine Cultures® 39 Fully sequenced microbes 45 Microbial technical resource 61 Instructions for rehydrating freeze-dried cultures 63 ATCC MTA All care has been taken in the compilation of the information contained in this catalogue. However, any applications for products are suggestions only, and LGC Standards makes no express or implied representations or warranties regarding the accuracy, content, completeness, or reliability of the information or any suggestions provided, and specifically disclaims any and all implied warranties with respect to the same, including without limitation any warranties of merchantability, fitness or suitability for particular purpose. About ATCC Mission ATCC is a global nonprofit bioresource centre and ATCC is an independent, private, nonprofit research organisation that provides biological biological resource centre (BRC) and research products, technical services and educational organisation. programmes to private industry, government and academic organisations. The mission of As a biological resource centre, ATCC ATCC is to acquire, authenticate, preserve, authenticates microorganisms and cell lines and develop and distribute biological materials, manages logistics of long-term preservation information, technology, intellectual
    [Show full text]
  • Mycoplasma Spermatophilum, a New Species Isolated from Human Spermatozoa and Cervix AURIOL C
    INTERNATIONALJOURNAL OF SYSTEMATICBACTERIOLOGY, Apr. 1991, p. 229-233 Vol. 41, No. 2 0020-7713/91/020229-05$02.oo/o Copyright 0 1991, International Union of Microbiological Societies Mycoplasma spermatophilum, a New Species Isolated from Human Spermatozoa and Cervix AURIOL C. HILL Medical Research Council Toxicology Unit, Woodmansterne Road, Carshalton, Surrey SM5 4EF, United Kingdom A mycoplasma isolated from human spermatozoa and a human cervix was shown to be serologically distinct from 98 previously recognized MycopEasma and AchoZepZasma spp. Six mycoplasma colonies were cloned and examined in detail for morphology, growth, and biochemical characteristics; five of these were from sperm samples and one was from a cervix. These strains were closely related and had the following properties: guanine-plus-cytosine content of 32 mol%, requirement for sterol, and anaerobic growth. Glucose was not metabolized, and arginine and urea were not hydrolyzed. Strain AH159 (= NCTC 11720) is the type strain of a new species, Mycoplasma spermatophilum. Twelve named Mycoplasma and Acholeplasma species colony isolated from each patient was cloned to produce a have been isolated from the respiratory or genital tracts of pure culture; this was done by initially filtering a broth humans (6). Mycoplasma buccale, Mycoplasma faucium, culture through a 220-nm-pore-size membrane filter, cultur- Mycoplasma lipophilum , Mycoplasma orale, Mycoplasma ing the filtrate on solid medium, transferring a single result- pneumoniae, and Mycoplasma salivarium are found almost ing colony to another agar plate, and inoculating the subse- exclusively in respiratory tracts. Mycoplasma fermentans quent growth into broth. This whole procedure was repeated has been found infrequently in urogenital tracts, while My- an additional four times; thus, the organisms were filter coplasma primatum, a species commonly present in nonhu- cloned five times (29).
    [Show full text]
  • United States Patent (19) 11 Patent Number: 5,824,479 Smida Et Al
    O USOO5824479A United States Patent (19) 11 Patent Number: 5,824,479 Smida et al. 45) Date of Patent: Oct. 20, 1998 54 INTER-LINE-PCR Chemical Abstract, vol. 120, 1994, p. 326. Abstract No. 20:976646, 75 Inventors: Jan Smida, Freising; Stefan Leibhard, Chemical Abstract, vol. 118, 1993, p. 562, Abstract No. Oberschleissheim; Ludwig Hieber, 118:56972r. Kirchheim; F. Eckardt-Schupp, Frothingham et al. A PCR-Based Method of Identifying Pfaffenhofen, all of Germany Species-Specific Repeated DNAs, BioTechniques, vol. 13. No. 2, 1992, pp. 210, 212. 213. 73) Assignee: Forschungszentrum fur, Umwelt und Furano et al., Amplification of the Ancient Murine Lx Family Gesundheit GmbH. Oberschleissheim, of Long Interspersed Repeated DNA Occurred During the Germany Murine Radiation, J. Mol. Evol., vol. 38 (1994), pp. 18-27. Pascale et al. The Evolution of Long Interspersed Repeated (21) Appl. No.: 646,809 DNA (L1, LINE 1) as Revealed by the Analysis of an Ancient Rodent Ll DNA Family, J. Mol. Evol. (1993), vol. 36 ppi (22 Filed: May 21, 1996 9-20. 30 Foreign Application Priority Data Gong et al., COMMUNICATION Identification of Region Specific Cosmid Clones by Hybrodization with Pooled May 22, 1995 DEI Germany ........................ 19518769.5 Alu-LINE Polymerase Chain Reaction Products of Yeast (51) int. Cl. ................... C12Q 1/68; C12P 19/34; Artificial Chromosone Clones, Methods in Mol. and Cell. CO7H 21/04 Biol. (1994), 4:269-272. (52) U.S. Cl. ........................... 435/6; 536/24.3: 536/22.1; Laten and Morris. SIRE-1, a long interspersed repetitive 435/91.2 DNA element from soybean with weak sequence similarity to (58) Field of Search .................................
    [Show full text]
  • Microbial Signatures of Oral Dysbiosis, Periodontitis and Edentulism 2 Revealed by Gene Meter Methodology 3 4 M
    bioRxiv preprint doi: https://doi.org/10.1101/070367; this version posted August 19, 2016. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 1 Microbial Signatures of Oral Dysbiosis, Periodontitis and Edentulism 2 Revealed by Gene Meter Methodology 3 4 M. Colby Hunter1, Alex E. Pozhitkov2, and Peter A. Noble#3 5 6 *Corresponding author, Peter A Noble, Email: [email protected] 7 8 Authors’ affiliations: 9 10 1. Program in Microbiology, Alabama State University, Montgomery, AL 36101 11 2. Department of Oral Health, University of Washington, Box 3574444, Seattle, Washington 12 98195-7444 Ph: 206-409-6664 13 3. Department of Periodontics, University of Washington, Box 3574444, Seattle, Washington 14 98195-7444 Ph: 206-409-6664 15 16 Authors’ emails: 17 18 Hunter: [email protected] 19 Pozhitkov: [email protected] 20 Noble: [email protected] 21 22 bioRxiv preprint doi: https://doi.org/10.1101/070367; this version posted August 19, 2016. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 23 ABSTRACT (N=305 WORDS) 24 25 Conceptual models suggest certain microorganisms (e.g., the red complex) are indicative of a specific 26 disease state (e.g., periodontitis); however, recent studies have questioned the validity of these models. 27 Here, the abundances of 500+ microbial species were determined in 16 patients with clinical signs of 28 one of the following oral conditions: periodontitis, established caries, edentulism, and oral health.
    [Show full text]
  • Moving Beyond Serovars
    ABSTRACT Title of Document: MOLECULAR AND BIOINFORMATICS APPROACHES TO REDEFINE OUR UNDERSTANDING OF UREAPLASMAS: MOVING BEYOND SEROVARS Vanya Paralanov, Doctor of Philosophy, 2014 Directed By: Prof. Jonathan Dinman, Cell Biology and Molecular Genetics, University of Maryland College Park Prof. John I. Glass, Synthetic Biology, J. Craig Venter Institute Ureaplasma parvum and Ureaplasma urealyticum are sexually transmitted, opportunistic pathogens of the human urogenital tract. There are 14 known serovars of the two species. For decades, it has been postulated that virulence is related to serotype specificity. Understanding of the role of ureaplasmas in human diseases has been thwarted due to two major barriers: (1) lack of suitable diagnostic tests and (2) lack of genetic manipulation tools for the creation of mutants to study the role of potential pathogenicity factors. To address the first barrier we developed real-time quantitative PCRs (RT-qPCR) for the reliable differentiation of the two species and 14 serovars. We typed 1,061 ureaplasma clinical isolates and observed about 40% of isolates to be genetic mosaics, arising from the recombination of multiple serovars. Furthermore, comparative genome analysis of the 14 serovars and 5 clinical isolates showed that the mba gene, used for serotyping ureaplasmas was part of a large, phase variable gene system, and some serovars shown to express different MBA proteins also encode mba genes associated with other serovars. Together these data suggests that differential pathogenicity and clinical outcome of an ureaplasmal infection is most likely due to the presence or absence of potential pathogenicity factors in individual ureaplasma clinical isolates and/or patient to patient differences in terms of autoimmunity and microbiome.
    [Show full text]
  • Rapid, Sensitive PCR-Based Detection of Mycoplasmas In
    APPLIED AND ENVIRONMENTAL MICROBIOLOGY, Mar. 1994, p. 953-959 Vol. 60, No. 3 0099-2240/94/$04.00 +0 Copyright © 1994, American Society for Microbiology Rapid, Sensitive PCR-Based Detection of Mycoplasmas in Simulated Samples of Animal Sera OLIVIER DUSSURGET AND DAISY ROULLAND-DUSSOIX* Laboratoire des Mycoplasnes, Institut Pasteur, 75724 Paris Cedex 15, Franice Received 2 August 1993/Accepted 17 December 1993 A fast and simple method to detect mycoplasmal contamination in simulated samples of animal sera by using a PCR was developed. The following five mycoplasma species that are major cell culture contaminants belonging to the class Mollicutes were investigated: Mycoplasma arginini, Acholeplasma laidlawii, Mycoplasma hyorhinis, Mycoplasma orale, and Mycoplasma fermentans. After a concentration step involving seeded sera, genus-specific primers were used to amplify a 717-bp DNA fragment within the 16S rRNA gene of mycoplasmas. In a second step, the universal PCR was followed by amplification of variable regions of the 16S rRNA gene by using species-specific primers, which allowed identification of contaminant mycoplasmas. With this method, 10 fg of purified DNA and 1 to 10 color-changing units of mycoplasmas could be detected. Since the sensitivity of the assay was increased 10-fold when the amplification products were hybridized with an internal mycoplasma-specific 32P-labelled oligonucleotide probe, a detection limit of 1 to 10 genome copies per PCR sample was obtained. This highly sensitive, specific, and simple assay may be a useful alternative to methods currently used to detect mycoplasmas in animal sera. Mycoplasmas (the trivial name for microorganisms belong- Like most cell cultures infected by mycoplasmas, serum infec- ing to the class Molliclutes) are the smallest self-replicating tions are rarely detected by visual inspection or light micros- bacteria.
    [Show full text]