DOCSLIB.ORG

Corynebacteriae

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Corynebacteriae Study of the properties of channel-forming proteins of the cell walls of different Corynebacteriae Dissertation zur Erlangung des naturwissenschaftlichen Doktorgrades der Fakultät für Biologie an der Bayerischen Julius-Maximilians-Universität Würzburg vorgelegt von Enrico Barth Würzburg Würzburg 2008 Eingereicht am: ...................................................................................... Prüfungskommission: Vorsitzender: Prof. Dr. Martin Müller, Dekan der Fakultät für Biologie 1. Gutachter: Prof. Dr. Dr. h.c. Roland Benz 2. Gutachter: Prof. Dr. Roy Gross Tag der mündlichen Prüfung: ................................................................ Promotionsurkunde ausgehändigt am: ................................................. Ich versichere, dass ich diese Dissertation selbständig angefertigt und keine anderen, als die von mir angegeben Quellen und Hilfsmittel benutzt habe. Die von mir vorgelegte Dissertation hat noch in keinem früheren Prüfungsverfahren in ähnlicher oder gleicher Form vorgelegen. Ich habe zu keinem früheren Zeitpunkt versucht, einen akademischen Grad zu erwerben. .......................................... Contents Contents................................................................................................................................ 9 Chapter 1 .............................................................................................................................. 15 1.1 Introduction ............................................................................................................. 15 1.2 Bacterial cell wall structures .................................................................................... 16 1.2.1 The cytoplasmic membrane ........................................................................................... 16 1.2.2 The periplasmic space ..................................................................................................... 17 1.2.3 The peptidoglycan layer .................................................................................................. 18 1.2.4 The cell wall of Gram-positive bacteria ....................................................................... 21 1.2.5 The cell wall of Gram-negative bacteria ...................................................................... 23 1.2.6 The cell wall of mycolic acid containing bacteria ....................................................... 26 1.2.6.1 The arabinogalactan moiety ........................................................................................... 29 1.2.6.2 The mycolic acid moiety................................................................................................. 30 1.2.6.3 Lipoarabinomannan ........................................................................................................ 31 1.2.6.4 Extractable lipids ............................................................................................................. 33 1.2.6.5 Porins penetrate the mycolic acid layer ........................................................................ 35 1.3 Aims of the work ...................................................................................................... 38 Chapter 2 ............................................................................................................................. 39 Corynebacterium diphtheriae: identification and characterization of a channel- forming protein in the cell wall ........................................................................................... 39 2.1 Summary .................................................................................................................. 39 2.2 Introduction ............................................................................................................. 40 2.3 Materials and Methods ............................................................................................ 42 2.3.1 Bacterial strains and growth conditions ....................................................................... 42 2.3.2 Construction of C. glutamicum strain ATCC 13032 ΔporHΔporA ............................. 42 2.3.3 Preparation of the cell wall, plasma membrane and cytosol fractions .................... 43 2.3.4 Isolation and purification of the channel-forming protein from the cell wall fraction .............................................................................................................................. 44 2.3.5 Digestion of the polypeptide ......................................................................................... 44 2.3.6 SDS-polyacrylamide gel electrophoresis ...................................................................... 44 2.3.7 Immunological techniques ............................................................................................. 45 2.3.7.1 Western blot analysis ...................................................................................................... 45 2.3.7.2 Enzyme-linked immunosorbent assay .. ...................................................................... 45 2.3.8 Polymerase chain reaction and construction of the expression plasmid ................ 46 2.3.9 Lipid bilayer experiments ............................................................................................... 47 9 2.4 Results ..................................................................................................................... 47 2.4.1 Isolation and purification of the channel-forming protein ........................................ 47 2.4.2 Interaction of the cell wall protein with lipid bilayer membranes ............................ 49 2.4.3 Single-channel analysis .................................................................................................... 49 2.4.4 Selectivity of the cell wall channel of C. diphtheriae ..................................................... 52 2.4.5 The cell wall channel of C. diphtheriae is voltage dependent ...................................... 53 2.4.6 Immunological detection of the channel-forming protein of C. diphtheriae............. 55 2.4.7 Identification of the gene coding for the cell wall channel of C. diphtheriae ATCC 11913 ..................................................................................................................... 56 2.4.8 Expression of CdPorA in C. glutamicum ATCC 13032 ΔporHΔporA and study of its channel-forming ability ............................................................................... 57 2.5 Discussion ............................................................................................................... 58 2.5.1 The cell wall of C. diphtheriae contains a channel-forming oligomer ........................ 58 2.5.2 The cell wall channel of C. diphtheriae is wide and water-filled and does not contain point charges ...................................................................................................... 59 2.5.3 The cell wall channel of C. diphtheriae shows immunological homology with CgPorA of C. glutamicum ................................................................................................. 60 2.5.4 Cloning and sequencing of cdporA................................................................................. 60 2.5.5 CdporA can be expressed in C. glutamicum and complements for CgPorA and CgPorH deficiency ........................................................................................................... 63 Chapter 3 ............................................................................................................................. 65 Study of the major cell wall channel of Corynebacterium glutamicum, Corynebacterium efficiens and Corynebacterium diphtheriae in dependence of PorH and PorA proteins ..................................................................................................... 65 3.1 Summary .................................................................................................................. 65 3.2 Introduction ............................................................................................................ 66 3.3 Materials and methods ............................................................................................ 68 3.3.1 Bacterial strains and growth conditions........................................................................ 68 3.3.2 Plasmids and DNA manipulations ................................................................................ 68 3.3.2.1 Cloning of C. glutamicum porins...................................................................................... 69 3.3.2.2 Cloning of C. efficiens porins ............................................................................................ 70 3.3.2.3 Cloning of C. diphtheriae porins ...................................................................................... 70 3.3.2.4 Cloning of the porin domain of C. callunae .................................................................. 73 3.3.3 Extraction of RNA .......................................................................................................... 73 3.3.4 DIG-labelled DNA probes ............................................................................................ 74 3.3.5 Northern blot analysis ..................................................................................................... 74 3.3.6 Reverse transcription ......................................................................................................
Load more

Recommended publications
  • Accuprobe Mycobacterium Avium Complex Culture
    non-hybridized and hybridized probe. The labeled DNA:RNA hybrids are measured in a Hologic luminometer. A positive result is a luminometer reading equal to or greater than the cut-off. A value below this cut-off is AccuProbe® a negative result. REAGENTS Note: For information on any hazard and precautionary statements that MYCOBACTERIUM AVIUM may be associated with reagents, refer to the Safety Data Sheet Library at www.hologic.com/sds. COMPLEX CULTURE Reagents for the ACCUPROBE MYCOBACTERIUM AVIUM COMPLEX IDENTIFICATION TEST CULTURE IDENTIFICATION TEST are provided in three separate reagent kits: INTENDED USE The ACCUPROBE MYCOBACTERIUM AVIUM COMPLEX CULTURE ACCUPROBE MYCOBACTERIUM AVIUM COMPLEX PROBE KIT IDENTIFICATION TEST is a rapid DNA probe test which utilizes the Probe Reagent. (4 x 5 tubes) technique of nucleic acid hybridization for the identification of Mycobacterium avium complex Mycobacterium avium complex (M. avium complex) isolated from culture. Lysing Reagent. (1 x 20 tubes) Glass beads and buffer SUMMARY AND EXPLANATION OF THE TEST Infections caused by members of the M. avium complex are the most ACCUPROBE CULTURE IDENTIFICATION REAGENT KIT common mycobacterial infections associated with AIDS and other Reagent 1 (Lysis Reagent). 1 x 10 mL immunocompromised patients (7,15). The incidence of M. avium buffered solution containing 0.04% sodium azide complex as a clinically significant pathogen in cases of chronic pulmonary disease is also increasing (8,17). Recently, several Reagent 2 (Hybridization Buffer). 1 x 10 mL laboratories have reported that the frequency of isolating M. avium buffered solution complex is equivalent to or greater than the frequency of isolating M.
    [Show full text]
  • Strategies for Cryo-Electron Tomography of the Mycobacterial Cell Envelope and Its Pore Proteins and Functional Studies of Porin Mspa from Mycobacterium Smegmatis
    Technische Universität München Max-Planck-Institut für Biochemie Abteilung für Molekulare Strukturbiologie Strategies for cryo-electron tomography of the mycobacterial cell envelope and its pore proteins and functional studies of porin MspA from Mycobacterium smegmatis Christian Werner Hoffmann Vollständiger Abdruck der von der Fakultät für Chemie der Technischen Universität München zur Erlangung des akademischen Grades eines Doktors der Naturwissenschaften genehmigten Dissertation. Vorsitzender: Univ. – Prof. Dr. J. Buchner Prüfer der Dissertation: 1. Hon. – Prof. Dr. W. Baumeister 2. Univ. – Prof. Dr. S. Weinkauf 3. Univ. – Prof. Dr. W. Liebl Die Dissertation wurde am 28.04.2010 bei der Technischen Universität München eingereicht und durch die Fakultät für Chemie am 14.07.2010 angenommen. TABLE OF CONTENTS A Table of contents A Table of contents .................................................................................................................... I B Abbreviations ........................................................................................................................ V C Zusammenfassung............................................................................................................ VIII D Summary ................................................................................................................................ X 1 Introduction ........................................................................................................................... 1 1.1 The genus Mycobacterium .............................................................................................
    [Show full text]
  • Expression and Regulation of the Porin Gene Mspa of Mycobacterium Smegmatis
    Expression and regulation of the porin gene mspA of Mycobacterium smegmatis Den Naturwissenschaftlichen Fakultäten der Friedrich-Alexander-Universität Erlangen-Nürnberg zur Erlangung des Doktorgrades vorgelegt von Dietmar Hillmann aus Nürnberg Als Dissertation genehmigt von den Naturwissenschaftlichen Fakultäten der Universität Erlangen-Nürnberg Tag der mündlichen Prüfung: 15.12.2006 Vorsitzender der Promotionskommission: Prof. Dr. E. Bänsch Erstberichterstatter: Prof. Dr. M. Niederweis Zweitberichterstatter: Prof. Dr. A. Burkovski Index Index 1 Zusammenfassung / Summary 1 2 Introduction 2 2.1 The genus Mycobacterium 2 2.1.1 Taxonomy 2 2.1.2 The architecture of the mycobacterial cell wall 3 2.2 Porins: Structure and function in gram-negative bacteria 5 2.3 Mycobacterial porins 7 2.4 Porin regulation 10 2.5 Expression of mspA of M. smegmatis 12 2.6 Scope of the thesis 13 3 Results 14 3.1 Screening system to monitor mycobacterial promoter activity 14 3.2 Transcriptional mechanisms affecting mspA expression 18 3.2.1 Identification of the mspA promoter 18 3.2.2 A very long upstream DNA element is required for full activity of pmspA 22 3.2.3 Influence of translation initiation signals of pmspA on lacZ expression 24 3.2.4 Influence of a distal DNA element on pmspA activation 25 3.2.5 Alignment of the 5’ regions of mspA, mspB, mspC and mspD 27 3.3 Post-transcriptional mechanisms affecting mspA expression 28 3.3.1 Detection of an antisense RNA to the mspA transcript 28 3.3.2 Secondary structure of the 5’ UTR of mspA 31 3.4 pH dependent mspA expression
    [Show full text]
  • Structure of the Mycobacterium Tuberculosis Ompatb Protein: a Model of an Oligomeric Channel in the Mycobacterial Cell Wall
    proteins STRUCTURE O FUNCTION O BIOINFORMATICS Structure of the Mycobacterium tuberculosis OmpATb protein: A model of an oligomeric channel in the mycobacterial cell wall Yinshan Yang,1,2 Daniel Auguin,1,2,3 Ste´phane Delbecq,4 Emilie Dumas,1,2 Ge´rard Molle,1,2 Virginie Molle,5 Christian Roumestand,1,2* and Nathalie Saint1,2 1 Centre de Biochimie Structurale, CNRS UMR 5048, Universite´ Montpellier 1 et 2, F34090 Montpellier, France 2 INSERM U554 F34090 Montpellier, France 3 INRA, USC2030 ‘Arbres et Re´ponses aux Contraintes Hydrique et Environnementales’ (ARCHE), F-45067 Orle´ans Cedex 02, France 4 Dynamique des Interactions Membranaires Normales et Pathologiques, CNRS UMR 5235, Universite´ Montpellier 2, F34095 Montpellier Cedex 5, France 5 Institut de Biologie et de Chimie des Prote´ines, Universite´ de Lyon, CNRS UMR 5086, F69367 Lyon, France ABSTRACT INTRODUCTION The pore-forming outer membrane protein The etiological agent of tuberculosis (TB), Mycobacterium OmpATb from Mycobacterium tuberculosis is a viru- tuberculosis, causing nearly 2 million deaths per year, is presently lence factor required for acid resistance in host one of the most important infectious agents implicated in mortal- phagosomes. In this study, we determined the 3D ity worldwide. TB has emerged as a major public health threat structure of OmpATb by NMR in solution. We because of a significant increase in multiple-drug-resistant TB and found that OmpATb is composed of two independ- synergism between human immunodeficiency virus and M. tuber- ent domains separated by a proline-rich hinge culosis infection.1,2 One of the principle problems in TB therapy region.
    [Show full text]
  • Actinomycesspp & Nocardiaspp
    PLUMB’S THERAPEUTICS BRIEF h PATHOGEN PROFILE h PEER REVIEWED Actinomyces spp & Nocardia spp J. Scott Weese, DVM, DVSc, DACVIM University of Guelph Actinomyces spp and Nocardia spp are members of class Acti- Nocardia spp nobacteria, which can cause opportunistic infections in dogs, The Nocardia genus contains more than 30 saprophytic species cats, and other species. Both can cause pyogranulomatous or that are widely, if not ubiquitously, disseminated in the environ- suppurative disease that is often slowly progressing and chal- ment. Disease occurs following inoculation of the bacterium into lenging to diagnose; however, some differences in organism tissue or via inhalation. and characteristics exist (Table 1, next page). The incidence of h Nocardia spp are regionally variable, with higher rates of disease caused by either is undefined and likely low. infection in dry, dusty, and windy regions (eg, southwestern United States, parts of Australia). Actinomyces spp h Nocardiosis is classically divided into 3 clinical forms: Many different species are present, and taxonomy continues to pulmonary, disseminated (systemic), and cutaneous/ change; some species previously known as Actinomyces have subcutaneous.11-15 been reclassified in other genera, such as Arcanobacterium spp • Clinical presentations are as would be expected with pul- and Trueperella spp. Regardless, disease aspects remain monary, systemic, or cutaneous infections. Pulmonary or unchanged. Commonly found as part of the oral, GI, and genital cutaneous disease can progress to disseminated disease. microbiotas,1-3 Actinomyces spp and related genera are of lim- ited virulence unless inoculated into tissue (eg, via bites, foreign Diagnosis bodies, or trauma). Early diagnosis may be missed.
    [Show full text]
  • Antibitoic Treatment for Tuberculosis Induces a Profound Dysbiosis of the Gut Microbiome That Persists Long After Therapy Is Completed
    ANTIBITOIC TREATMENT FOR TUBERCULOSIS INDUCES A PROFOUND DYSBIOSIS OF THE GUT MICROBIOME THAT PERSISTS LONG AFTER THERAPY IS COMPLETED A Thesis Presented to the Faculty of the Weill Cornell Graduate School of Medical Sciences in Partial Fulfillment of the Requirements for the Degree of Masters of Science by Matthew F. Wipperman May 2017 © 2017 Matthew F. Wipperman ABSTRACT Mycobacterium tuberculosis, the cause of Tuberculosis (TB), infects one third of the world’s population and causes substantial mortality worldwide. In its shortest format, treatment of drug sensitive TB requires six months of multidrug therapy with a mixture of broad spectrum and mycobacterial specific antibiotics, and treatment of multidrug resistant TB is much longer. The widespread use of this regimen worldwide makes this one the largest exposures of humans to antimicrobials, yet the effects of antimycobacterial agents on intestinal microbiome composition and long term stability are unknown. We compared the microbiome composition, assessed by both 16S rDNA and metagenomic DNA sequencing, of Haitian TB cases during antimycobacterial treatment and following cure by 6 months of TB therapy. TB treatment does not perturb overall diversity, but nonetheless dramatically depletes multiple immunologically significant commensal bacteria. The perturbation by TB therapy lasts at least 1.5 years after completion of treatment, indicating that the effects of TB treatment are long lasting and perhaps permanent. These results demonstrate that TB treatment has dramatic and durable effects on the intestinal microbiome and highlight unexpected extreme consequences of treatment for the world’s most common infection on human ecology. BIOGRAPHICAL SKETCH NAME POSITION TITLE Wipperman, Matthew Frederick Postdoctoral Researcher at eRA COMMONS USER NAME Memorial Sloan Kettering Cancer Center MFWIPPERMAN DEGREE INSTITUTION AND (if MM/YY FIELD OF STUDY LOCATION applicable) Franklin & Marshall College B.A.
    [Show full text]
  • Evaluation Ofapi Coryne System for Identifying Coryneform Bacteria
    756 Y Clin Pathol 1994;47:756-759 Evaluation of API Coryne system for identifying coryneform bacteria J Clin Pathol: first published as 10.1136/jcp.47.8.756 on 1 August 1994. Downloaded from A Soto, J Zapardiel, F Soriano Abstract that are aerobe or facultatively aerobe, non- Aim-To identify rapidly and accurately spore forming organisms of the following gen- coryneform bacteria, using a commercial era: Corynebacterium, Listeria, Actinomyces, strip system. Arcanobacterium, Erysipelothrix, Oerskovia, Methods-Ninety eight strains of Cory- Brevibacterium and Rhodococcus. It also per- nebacterium species and 62 additional mits the identification of Gardnerella vaginalis strains belonging to genera Erysipelorix, which often has a diphtheroid appearance and Oerskovia, Rhodococcus, Actinomyces, a variable Gram stain. Archanobacterium, Gardnerella and We studied 160 organisms in total from dif- Listeria were studied. Bacteria were ferent species of the Corynebacterium genus, as identified using conventional biochemi- well as from other morphological related gen- cal tests and a commercial system (API- era or groups, some of them not included in Coryne, BioMerieux, France). Fresh rab- the API Coryne database. bit serum was added to fermentation tubes for Gardnerella vaginalis isolates. Results-One hundred and five out ofthe Methods 160 (65.7%) organisms studied were cor- The study was carried out on Gram positive rectly and completely identified by the bacilli belonging to the genera Coryne- API Coryne system. Thirty five (21.8%) bacterium, Erysipelothrix, Oerskovia, Rhodococcus, more were correctly identified with addi- Actinomyces, Arcanobacterium, Gardnerella and tional tests. Seventeen (10-6%) organisms Listeria included in the API Coryne database were not identified by the system and (table 1).
    [Show full text]
  • Mapping Interactions of Microbial Metabolites and Human Receptors
    bioRxiv preprint doi: https://doi.org/10.1101/614537; this version posted May 2, 2019. 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. Classification: Biological Sciences - Microbiology Title: Mapping interactions of microbial metabolites and human receptors Authors: 1Dominic A. Colosimo, 1Jeffrey A. Kohn, 1Peter M. Luo, 3Sun M. Han, 2AmanDa J. PickarD, 2Arka Rao, 2Justin R. Cross, 3Louis J. Cohen, 1Sean F. BraDy* Author affiliation: 1Laboratory of Genetically EncoDeD Small Molecules, The Rockefeller University, 1230 York Avenue, New York City, NY 10065. 2DonalD B. anD Catherine C. Marron Cancer Metabolism Center, Memorial Sloan Kettering Cancer Center, New York City NY 10065, USA. 3Division of Gastroenterology, Department of MeDicine, Icahn School of MeDicine at Mount Sinai, New York City, NY 10029, USA. *Corresponding Authors Sean F. BraDy Contact: Laboratory of Genetically EncoDeD Small Molecules The Rockefeller University 1230 York Avenue New York, NY 10065 Phone: 212-327-8280 Fax: 212-327-8281 Email: sbraDy@rockefeller.eDu Acknowledgements: All bacterial strains were generously proviDeD by Daniel MuciDa. High-resolution mass spectrometry of purifieD compounDs was performeD by Rockefeller University Proteomics Core. We are grateful to C. Fermin, E. Vazquez, anD G. Escano in the Precision Immunology Institute at the Icahn School of MeDicine at Mount Sinai (PrIISM) Gnotobiotic facility anD Microbiome Translational Center for their help with gnotobiotic experiments. FunDing was proviDeD by the Bill anD MelinDa Gates FounDation (OPP1168674) anD the National Institutes of Health (5R01AT009562–02). Keywords: primary metabolites, human microbiome, G-protein coupleD receptors Author Contributions: D.A.C., L.J.C.
    [Show full text]
  • Lesson 20. Mycobacterium
    Mycobacterium MODULE Microbiology 20 MYCOBACTERIUM Notes 20.1 INTRODUCTION Mycobacterium are slender rods that sometimes show branching filamentous forms resembling fungal mycelium. In liquid cultures they form a mould-like pellicle. Hence the name ‘mycobacteria’, meaning fungus like bacteria. They do not stain readily, but once stained, resist decolourisation with dilute mineral acids. Hence they are called ‘Acid fast bacilli’. They are aerobic, nonmotile, noncapsulated and nonsporing. OBJECTIVES After reading this lesson, you will be able to: z describe the morphology of Mycobacterium tuberculosis & M. leprae z describe the characteristics of Mycobacterium tuberculosis & M. leprae z explain about pathogenesis of Mycobacterium tuberculosis & M. leprae z explain the laboratory diagnosis Mycobacterium tuberculosis & M. leprae The first member of this genus to be identified was Lepra bacillus discovered by Hansen. Koch (1882) isolated the mammalian tubercle bacillus and proved its causative role in tuberculosis. In humans tuberculosis is caused by mycobacterium tuberculosis and also by bovine type called Mycobacterium bovis. The second human pathogenic mycobacterium is the lepra bacillus causing Leprosy. The third group of mycobacterium is a mixed group from varied sources like birds, cold-blooded and warm blooded animals, from skin ulcers, soil, water and other environmental sources. They are called as atypical mycobacteria. They are opportunistic pathogens and can cause many types of diseases. MICROBIOLOGY 203 MODULE Mycobacterium Microbiology 20.2 MYCOBACTERIUM TUBERCULOSIS Morphology M tuberculosis is a straight or slightly curved rod, about 3 X 0.3 µm in size, occurring singly, in pairs or as small clumps. M bovis is usually straighter, shorter and stouter. Tubercle bacilli have been described as Gram positive, even though after Notes staining with basic dyes they resist decolourisation by alcohol even without the effect of iodine.
    [Show full text]
  • Diarylcoumarins Inhibit Mycolic Acid Biosynthesis and Kill Mycobacterium Tuberculosis by Targeting Fadd32
    Diarylcoumarins inhibit mycolic acid biosynthesis and kill Mycobacterium tuberculosis by targeting FadD32 Sarah A. Stanleya,b,c, Tomohiko Kawatea,b,c, Noriaki Iwasea,b,c, Motohisa Shimizua,b,c, Anne E. Clatworthya,b,c, Edward Kazyanskayaa, James C. Sacchettinid, Thomas R. Ioergere, Noman A. Siddiqif, Shoko Minamif, John A. Aquadroa, Sarah Schmidt Granta,b,c, Eric J. Rubinf, and Deborah T. Hunga,b,c,1 aInfectious Disease Initiative, Broad Institute of Harvard and MIT, Cambridge, MA 02142; bDepartment of Molecular Biology, Massachusetts General Hospital, Boston, MA 02114; cDepartment of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02115; Departments of dBiochemistry and Biophysics and eComputer Science, Texas A&M University, College Station, TX 77843; and fDepartment of Immunology and Infectious Diseases, Harvard School of Public Health, Boston, MA 02115 Edited* by John J. Mekalanos, Harvard Medical School, Boston, MA, and approved May 30, 2013 (received for review February 1, 2013) Infection with the bacterial pathogen Mycobacterium tuberculosis not yet in clinical trials, include Benzothiazinones that target imposes an enormous burden on global public health. New anti- decaprenylphosphoryl-β-d-ribose 2′-epimerase (DprE1) (2) and biotics are urgently needed to combat the global tuberculosis inhibitors of malate synthase, a glyoxylate shunt enzyme (10). In pandemic; however, the development of new small molecules is addition to their potential as drug candidates, these molecules hindered by a lack of validated drug targets. Here, we describe the are significant for having facilitated the identification of novel identification of a 4,6-diaryl-5,7-dimethyl coumarin series that kills targets for further efforts geared toward drug discovery.
    [Show full text]
  • Development of a SYBR Green Multiplex Real Time PCR For
    Development of a SYBR Green… Safar Ali. A. et al 241 ORIGINAL ARTICLE Development of a SYBR Green Multiplex Real Time PCR for Simultaneous Detection of Mycobacterium Tuberculosis and Nocardia Asteroides in Respiratory Samples Safar Ali Alizadeh1*, Amir Javadi2, Jalal Mardaneh3,4, Neda Nasirian5, Sajjad Alizadeh6, Maryam Mohammadbeigi7, Siamak Heidarzadeh8 5Department of pathology, School of Medicine, Qazvin University of Medical Sciences, OPEN ACCESS Qazvin, Iran. 6Medical Doctor, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran. Citation: Safar Ali Alizadeh, Amir 7Department of Microbiology and Immunology, School of Medicine, Qazvin University of Javadi, Jalal Mardaneh, Neda Nasirian, Medical Sciences, Qazvin, Iran. Sajjad Alizadeh, Maryam 8Department of Microbiology and Virology, School of Medicine, Zanjan University of Mohammadbeigi, Siamak Heidarzadeh. Medical Sciences, Zanjan, Iran. Development of an SYBR Green *Email: alizadeh120@yahoo.com Multiplex Real Time PCR for Simultaneous Detection of ABSTRACT Mycobacterium Tuberculosis and Nocardia Asteroides in Respiratory Samples. Ethiop J Health Sci. 2021; BACKGROUND፡ Nocardia asteroides and Mycobacterium 31(2):241 tuberculosis are worldwide-distributed bacteria. These infectious doi:http://dx.doi.org/10.4314/ejhs.v31i2.6 Received: October 27, 2020 agents can cause many infections in humans, especially in Accepted: November 23, 2020 immunocompromised individuals. Pulmonary infections are more Published: March 1, 2021 common and have similar clinical symptoms. Proper diagnosis Copyright : © 2021 Safar A.A.., et al. This is an open access article distributed and treatment of these patients are important for accurate under the terms of the Creative treatment and could be lifesaving. Commons Attribution License, which permits unrestricted use, distribution, METHODS: In this study, a multiplex real-time PCR assay was and reproduction in any medium, established for the simultaneous detection of the N.
    [Show full text]
  • Neisseria Obligate Aerobe Usmle
    Neisseria Obligate Aerobe Usmle Unreverent Steward coruscate very effectually while Olin remains unpitied and asclepiadaceous. Antiphrastic Remus rejoiced reportedly or predestining huffishly when Mohammed is brainier. How Latin is Aubert when genitival and woozier Barney gritted some dominee? Journal will cause a handy way down the obligate aerobe bacteria are notorious smallpox virus vaccine development Mechanism and their use clinical features is untreated, confirmed by tsetse fly, david i speak about viruses. By neisseria gonorrhea. Chlamydiae are obligate aerobes. Progresses rapidly and include biochemical foundations of obligate aerobe? Nearby is poorly immunogenic in development caused by hermann eibel. Which one vial should be classified as early sign in order shipped on. The bacterium can be monitored regularly for national microbiology laboratory screening after being affected most often causes no. Like receptor protein structures or conjunctival infections that binds siderophores. The obligate aerobes can improve treatment for misconfigured or diploid cells with their own atp? Gonococcal infections are obligate aerobe? Intracellular pathogen releases verotoxin, which is less important microorganisms is a mechanism of microbiology, give aspirin or disseminated infections. Dna into facebook confirmed results on their fe transport systems used for diagnostic testing are out fermentation for her unfailing devotion. The following are modeled to another bacterium, ascending gonococcal infections, and liver or with alkaline urine sample for fe can be more than the four days of all treatment. Key differences between cells may be visualized by entrance of electrical signals skin flora. Shigella sonnei with lecturio offers and decide how do stick or jump right lobe diagnosis. Recognition proteins cell and neisseria meningitidis, motor or stored as confirmation with symptoms.
    [Show full text]