DOCSLIB.ORG

Rapid Detection and Differentiation of Mycobacterial Species Using a Multiplex PCR System

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Rapid Detection and Differentiation of Mycobacterial Species Using a Multiplex PCR System Revista da Sociedade Brasileira de Medicina Tropical 46(4):447-452, Jul-Aug, 2013 http://dx.doi.org/10.1590/0037-8682-0097-2013 MCaseajor ReportArticle Rapid detection and differentiation of mycobacterial species using a multiplex PCR system Andrea Santos Lima[1], Rafael Silva Duarte[2], Lílian Maria Lapa Montenegro[1] and Haiana Charifker Schindler[1] [1]. Departamento de Imunologia, Centro de Pesquisas Aggeu Magalhães, Fundação Oswaldo Cruz, Recife, PE. [2]. Instituto de Microbiologia, Universidade Federal do Rio Janeiro, Rio de Janeiro, RJ. ABSTRACT Introduction: The early diagnosis of mycobacterial infections is a critical step for initiating treatment and curing the patient. Molecular analytical methods have led to considerable improvements in the speed and accuracy of mycobacteria detection. Methods: The purpose of this study was to evaluate a multiplex polymerase chain reaction system using mycobacterial strains as an auxiliary tool in the differential diagnosis of tuberculosis and diseases caused by nontuberculous mycobacteria (NTM) Results: Forty mycobacterial strains isolated from pulmonary and extrapulmonary origin specimens from 37 patients diagnosed with tuberculosis were processed. Using phenotypic and biochemical characteristics of the 40 mycobacteria isolated in LJ medium, 57.5% (n=23) were characterized as the Mycobacterium tuberculosis complex (MTBC) and 20% (n=8) as nontuberculous mycobacteria (NTM), with 22.5% (n=9) of the results being inconclusive. When the results of the phenotypic and biochemical tests in 30 strains of mycobacteria were compared with the results of the multiplex PCR, there was 100% concordance in the identifi cation of the MTBC and NTM species, respectively. A total of 32.5% (n=13) of the samples in multiplex PCR exhibited a molecular pattern consistent with NTM, thus disagreeing with the fi nal diagnosis from the attending physician. Conclusions: Multiplex PCR can be used as a differential method for determining TB infections caused by NTM a valuable tool in reducing the time necessary to make clinical diagnoses and begin treatment. It is also useful for identifying species that were previously not identifi able using conventional biochemical and phenotypic techniques. Keywords: Mycobacterium tuberculosis complex. Nontuberculous mycobacteria. Diagnosis. Polymerase chain reaction. Multiplex. guided by video, where the instruments are subjected to INTRODUCTION high levels of sterilization in glutaraldehyde solution. From 1998 to 2009, there were 2,520 reported cases of post- The ability of nontuberculous mycobacteria (NTM) to cause surgical infections related to rapidly growing mycobacteria, disease is clearly described in the literature, and its importance is including Mycobacterium fortuitum, Mycobacterium chelonae, increasing progressively, with an increasing amount of various M. abscessus and Mycobacterium massiliense, which were species being cultured and isolated in laboratories1-3. distributed predominantly in private hospitals throughout In Brazil, the NTM species most frequently associated with the country. Cases have been reported in 23 states of Brazil, 6 lung disease are Mycobacterium kasasii and Mycobacterium with 97.8% of the cases concentrated within 10 states . The avium. Other species, such as Mycobacterium xenopi, diseases caused by NTMs are not compulsorily notifi able, Mycobacterium malmoense, Mycobacterium lentiflavum, except where the infection occurred during surgical procedures; Mycobacterium abscessus and Mycobacterium szulgai, have however, several studies in Brazil have noted isolated strains of occasionally been isolated in cultures4.5. In 2011 the National nontuberculous mycobacteria in pulmonary and extrapulmonary 4,7,8. Agency for Sanitary Surveillance (ANVISA) published the clinical samples from patients with suspected tuberculosis number of cases of infections by rapidly growing mycobacteria The differential diagnosis between tuberculosis (TB) and (RGMs) obtained in the last eleven years in Brazil. diseases caused by NTMs is of great importance because Data also suggest that this disease occurs in subjects the epidemiology, treatment and prognosis are different. undergoing invasive procedures, mostly surgical cases Conventional diagnostics present numerous diffi culties as clinical symptoms are generally similar and non-specifi c smear testing has limited sensitivity and cannot differentiate the species of mycobacteria. a o Address to: Dr Andrea Santos Lima. Dept de Imunologia/CPqAM/FIOCRUZ. For many years, the identifi cation of mycobacteria was Av. Moraes Rego s/n, Cidade Universitária, 50670-420 Recife, PE, Brasil. undertaken by gathering phenotypic results and biochemical Phone: 55 81 2101-2569; Fax: 55 81 3453-1911 9,10 e-mail: [email protected] test results of isolated species in culture medium . In the last Received 6 May 2013 20 or 30 years, with the increasing need to identify a greater Accepted 22 July 2013 number of species and also for early diagnosis, new methods 447 Lima AS et al - Differentiation of mycobacterial species have been developed to enable rapid and accurate identifi cation from skin lesion aspiration, 3 (7.5%) from pleural fl uid (PL), of mycobacterial species10-12. In Brazil, the PRA-hsp65 1 (2.5%) from a node biopsy and 1 (2.5%) from a bone biopsy. technique proposed by Telenti et al.13, and Devallois et al.14 Clinical specimens were analyzed by bacilloscopy, culture, has enabled the identifi cation of several species of NTM, the multiplex PCR and polymerase chain reaction restriction results of which correlated well with those from biochemical enzyme analysis (PRA-hsp65) to identify and differentiate identification13-17. Beyond the molecular techniques, the mycobacterial species as described below. sequencing of the specifi c genes rpoB and hsp65 has become Smear microscopy the gold standard for identifying mycobacteria18,19. Because of its high discriminatory power, this tool has been used by The presence of acid fast bacilli in different clinical several groups in epidemiological studies to identify the species specimens was determined by Ziehl-Neelsen staining (Stewart, involved in human diseases and in outbreaks, and also for 1953) in the Public Health Laboratory of Recife (Unifi ed taxonomic reclassifi cation20,21. Health System - SUS), in accordance with the guidelines of Multiplex polymerase chain reaction (PCR) is a fast tool the Brazilian Ministry of Health29. that allows the simultaneous amplifi cation of more than one Specimen processing sequence of target deoxyribonucleic acid (DNA) in a single reaction, saving time and reagents22-24. This system, which In total, of 1-5 ml of various clinical specimens (except for amplifies two or three different targets, can differentiate the sterile samples collected) was processed using the modifi ed M. tuberculosis from the NTMs25,26. This molecular approach Petroff method in 4% NaOH29,30. is used in identifying and differentiating microbes because it is Culture able to supply a simple fi ngerprint of certain bacterial groups when compared to the standard profi les of referent strains. The Cultures were performed in a Lowenstein-Jensen medium29,31 correct choice of target sequences in the genome is one of the and mycobacterium species were identifi ed using the following key criteria for detecting and identifying mycobacteria by PCR27. biochemical tests: selective inhibition by para-nitrobenzoic Faced with the need to differentiate tuberculosis from other (PNB) acid and thiophene-2-carboxylic hydrazide (TCH) 29 mycobacteria, the aim of this study was to evaluate a system acid, niacin accumulation and heat-stable catalase at 68°C . based on multiplex PCR, already optimized by Poroca et al.26, The cultures analyzed using the biochemical tests in this study for identifying mycobacterial species isolated from different were considered as references for comparison with the results clinical samples from patients diagnosed with pulmonary and obtained by multiplex PCR and PRA-hsp65. extrapulmonary tuberculosis. This test, in addition to contributing DNA extraction to a rapid and accurate differential diagnosis between tuberculosis and other mycobacteria, will certainly assist in the early and DNA was extracted and purifi ed from reference strains of appropriate therapeutic management of the patient. Mycobacterium tuberculosis (H37Rv). In all, 40 species of mycobacteria were isolated on a Lowenstein-Jensen medium from clinical samples obtained from 37 patients diagnosed METHODS with pulmonary and extrapulmonary tuberculosis using the conventional method described by Sambrook et al. using a Study population mixture of solvents and phenol chloroform32. Thirty-seven male and female patients over the age of Polymerase chain reaction and restriction 12 years with pulmonary and extrapulmonary tuberculosis enzyme analysis diagnoses, who were referred to public clinics in the The polymerase chain reaction and restriction enzyme metropolitan region of Recife of northeastern Brazil between analysis (PRA-hsp65) were performed using the techniques March 2008 and December 2009, were chosen for this study. described in Telenti et al.13 The identifi cation was determined The types of biological samples collected were chosen based by comparing the sizes of the fragments with the algorithm on the different clinical forms of the disease. The diagnosis of described in the PRASITE site (http://app.chuv.ch/prasite/ tuberculosis was made by the attending physician in accordance index.html)17. with the standards
Load more

Recommended publications
  • Mycobacterium Arupense Among the Isolates of Non-Tuberculous Mycobacteria from Human, Animal and Environmental Samples
    Veterinarni Medicina, 55, 2010 (8): 369–376 Original Paper Mycobacterium arupense among the isolates of non-tuberculous mycobacteria from human, animal and environmental samples M. Slany1, J. Svobodova2, A. Ettlova3, I. Slana1, V. Mrlik1, I. Pavlik1 1Veterinary Research Institute, Brno, Czech Republic 2Regional Institute of Public Health, Brno, Czech Republic 3BioPlus, s.r.o., Brno, Czech Republic ABSTRACT: Mycobacterium arupense is a non-tuberculous, potentially pathogenic species rarely isolated from humans. The aim of the study was to ascertain the spectrum of non-tuberculous mycobacteria within 271 sequenced mycobacterial isolates not belonging to M. tuberculosis and M. avium complexes. Isolates were collected between 2004 and 2009 in the Czech Republic and were examined within the framework of ecological studies carried out in animal populations infected with mycobacteria. A total of thirty-three mycobacterial species were identified. This report describes the isolation of M. arupense from the sputum of three human patients and seven different animal and environmental samples collected in the last six years in the Czech Republic: one isolate from leftover refrigerated organic dog food, two isolates from urine and clay collected from an okapi (Okapia johnstoni) and antelope bongo (Tragelaphus eurycerus) enclosure in a zoological garden, one isolate from the soil in an eagle’s nest (Haliaeetus albicilla) band two isolates from two common vole (Microtus arvalis) livers from one cattle farm. All isolates were identified by biochemical tests, morphology and 16S rDNA sequencing. Also, retrospective screening for M. arupense occurrence within the collected isolates is presented. Keywords: 16S rDNA sequencing; non-tuberculous mycobacteria; ecology Non-tuberculous mycobacteria (NTM) are ubiq- According to the commonly used Runyon clas- uitous in the environment and are responsible for sification scheme, NTM are categorized by growth several diseases in humans and/or animals known rate and pigmentation.
    [Show full text]
  • Epidemiology of Infection by Nontuberculous Mycobacteria JOSEPH O
    CLINICAL MICROBIOLOGY REVIEWS, Apr. 1996, p. 177–215 Vol. 9, No. 2 0893-8512/96/$04.0010 Copyright q 1996, American Society for Microbiology Epidemiology of Infection by Nontuberculous Mycobacteria JOSEPH O. FALKINHAM III* Department of Biology, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24061-0406 INTRODUCTION .......................................................................................................................................................178 The Nontuberculous Mycobacteria.......................................................................................................................178 Nontuberculous Mycobacterial Disease before the AIDS Epidemic................................................................179 Nontuberculous Mycobacterial Disease after the AIDS Epidemic ..................................................................179 Current Trends in the Epidemiology of Nontuberculous Mycobacterial Disease .........................................179 Chemotherapy of Nontuberculous Mycobacterial Infections............................................................................180 Detection, Recovery, and Identification of Nontuberculous Mycobacteria.....................................................181 Ecology and Physiology of Nontuberculous Mycobacteria................................................................................181 Virulence of Nontuberculous Mycobacteria ........................................................................................................182
    [Show full text]
  • Sneha Bowalekar and Rahul Gadkari.Pdf
    Int.J.Curr.Microbiol.App.Sci (2021) 10(02): 476-482 International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 10 Number 02 (2021) Journal homepage: http://www.ijcmas.com Case Study https://doi.org/10.20546/ijcmas.2021.1002.056 A Rare Case of Olecranon Bursitis caused by Mycobacterium abscessus Treated with Surgical Intervention Sneha Bowalekar* and Rahul Gadkari 1Consultant Microbiologist, Dr. Jariwala Laboratory, 1st floor, Rasraj Heights, Rokadia Lane, Near Gokul Hotel, Boriwali West, Mumbai, Maharashtra, India 2Consultant Orthopedic Surgeon, Sailee Hospital and Diagnostic Centre, Prathamesh Horizon, New Link Road, Boriwali West, Mumbai, Maharashtra, India *Corresponding author ABSTRACT K e yw or ds The present case of Mycobacterium abscessus olecranon bursitis in an M. abscessus , immunocompetant male treated with surgical drainage without requirement of Olecranon bursitis, antimycobacterial therapy. Nontuberculous mycobacteria are widely listed as Immunocompetant an infective cause of olecranon bursitis and other skin and soft tissue Article Info infections. Direct inoculation of the organism during trauma leads to musculoskeletal infections in particular. We could not find any other case Accepted: report of olecranon bursitis caused by M. abscessus in the literature. This case 07 January 2021 extends the view about nontuberculous mycobacterial infections in Available Online: 10 February 2021 immunocometant adults with co-morbid condition such as diabetes. Introduction olecranon bursitis caused by NTM identified as Mycobacterium abscessus. The infection Nontuberculous mycobacteria (NTM) are was treated with surgical drainage of increasingly being reported as cause of olecranon bursa fluid. Clinical follow up of infections in immunocompromised as well as patient was uneventful. immunocompetant patients.
    [Show full text]
  • Clinical and Epidemiological Features
    P0506 Paper Poster Session III Nontuberculous mycobacteria Nontuberculous mycobacteria in a third level hospital in Spain: clinical and epidemiological features G. Barbeito Castiñeiras1, M. Otero1, L. Ferreiro1, R. Trastoy1, J.J. Costa1, V. Tuñez1, M.L. Pérez del Molino1 1Clinical Microbiology Department- Complexo Hospitalario Universitario de Santiago de Compostela, Santiago de Compostela, Spain INTRODUCTION In the last few years, we have been attending to an increasing number of isolations of non-tuberculous mycobacteria (NTM) in the health area of Santiago de Compostela (458.759 inhabitants). Our objective is to study the epidemiology of those infections caused by NTM, their associated factors and their clinical significance. METHOD Retrospective study of NTM isolations carried out from 2005 to 2013. Data sources: Microbiology Information System (OpenLab) and the electronic clinical history of Galicia (IANUS). Statistical analysis: SPSSv.20. Microbiological techniques: auramine staining, and the growth in liquid media (MGIT, Bactec 960, Becton Dickinson) 45 days and solid culture of Coletsos ® 8 weeks. Identification:phenotypic and genotypic methods: GenoType®Mycobacterium CM/AS (Hain Lifescience). For diagnosis, the criteria from the American Thoracic Society / Infectious Diseases Society of America (ATS/IDSA) 2007 were applied and the revision of the clinical history was used for the evaluation of clinical significance. RESULTS During those 9 years of study, a total of 456 strains were aisolated (Mycobacterium avium complex 34,65%, Mycobacterium intracellulare 20,83%, Mycobacterium xenopi 11,84%, Mycobacterium abscessus 9,21%, others 23,47%), concerning 212 patients. 91 patients fulfilled the NTM disease criteria of the ATS/IDSA (19,96%). The average age was 61 (range 1-89), 61,54% were male.
    [Show full text]
  • Nontuberculous Mycobacteria in Respiratory Samples from Patients with Pulmonary Tuberculosis in the State of Rondônia, Brazil
    Mem Inst Oswaldo Cruz, Rio de Janeiro, Vol. 108(4): 457-462, June 2013 457 Nontuberculous mycobacteria in respiratory samples from patients with pulmonary tuberculosis in the state of Rondônia, Brazil Cleoni Alves Mendes de Lima1,2/+, Harrison Magdinier Gomes3, Maraníbia Aparecida Cardoso Oelemann3, Jesus Pais Ramos4, Paulo Cezar Caldas4, Carlos Eduardo Dias Campos4, Márcia Aparecida da Silva Pereira3, Fátima Fandinho Onofre Montes4, Maria do Socorro Calixto de Oliveira1, Philip Noel Suffys3, Maria Manuela da Fonseca Moura1 1Centro Interdepartamental de Biologia Experimental e Biotecnologia, Universidade Federal de Rondônia, Porto Velho, RO, Brasil 2Laboratório Central de Saúde Pública de Rondônia, Porto Velho, RO, Brasil 3Laboratório de Biologia Molecular Aplicada a Micobactérias, Instituto Oswaldo Cruz 4Centro de Referência Professor Hélio Fraga, Escola Nacional de Saúde Pública-Fiocruz, Rio de Janeiro, RJ, Brasil The main cause of pulmonary tuberculosis (TB) is infection with Mycobacterium tuberculosis (MTB). We aimed to evaluate the contribution of nontuberculous mycobacteria (NTM) to pulmonary disease in patients from the state of Rondônia using respiratory samples and epidemiological data from TB cases. Mycobacterium isolates were identified using a combination of conventional tests, polymerase chain reaction-based restriction enzyme analysis of hsp65 gene and hsp65 gene sequencing. Among the 1,812 cases suspected of having pulmonary TB, 444 yielded bacterial cultures, including 369 cases positive for MTB and 75 cases positive for NTM. Within the latter group, 14 species were identified as Mycobacterium abscessus, Mycobacterium avium, Mycobacterium fortuitum, Myco- bacterium intracellulare, Mycobacterium gilvum, Mycobacterium gordonae, Mycobacterium asiaticum, Mycobac- terium tusciae, Mycobacterium porcinum, Mycobacterium novocastrense, Mycobacterium simiae, Mycobacterium szulgai, Mycobacterium phlei and Mycobacterium holsaticum and 13 isolates could not be identified at the species level.
    [Show full text]
  • Mycobacterium Avium Complex Olecranon Bursitis Resolves
    IDCases 2 (2015) 59–62 Contents lists available at ScienceDirect IDCases jo urnal homepage: www.elsevier.com/locate/idcr Case Report Mycobacterium avium complex olecranon bursitis resolves without antimicrobials or surgical intervention: A case report and review of the literature a, b a Selene Working *, Andrew Tyser , Dana Levy a Division of Infectious Diseases, University of Utah School of Medicine, 30 N 1900 E, Room 4B319, Salt Lake City, UT 84132, United States b Department of Orthopaedics, University Orthopaedic Center, 590 Wakara Way, Salt Lake City, UT 84108, United States A R T I C L E I N F O A B S T R A C T Article history: Introduction: Nontuberculous mycobacteria are an uncommon cause of septic olecranon bursitis, though Received 26 March 2015 cases have increasingly been described in both immunocompromised and immunocompetent hosts. Received in revised form 2 April 2015 Guidelines recommend a combination of surgical resection and antimicrobials for treatment. This case is Accepted 5 April 2015 the first reported case of nontuberculous mycobacterial olecranon bursitis that resolved without medical or surgical intervention. Keywords: Case presentation: A 67-year-old female developed a painless, fluctuant swelling of the olecranon bursa Nontuberculous mycobacteria following blunt trauma to the elbow. Due to persistent bursal swelling, she underwent three separate Olecranon bursitis therapeutic bursal aspirations, two involving intrabursal steroid injection. After the third aspiration, the Mycobacterium avium complex bursa became erythematous and severely swollen, and bursal fluid grew Mycobacterium avium complex. Triple-drug antimycobacterial therapy was initiated, but discontinued abruptly due to a rash. Surgery was not performed.
    [Show full text]
  • Mixed Cutaneous Infection Caused by Mycobacterium Szulgai and Mycobacterium Intermedium in a Healthy Adult Female: a Rare Case Report
    Hindawi Publishing Corporation Case Reports in Dermatological Medicine Volume 2015, Article ID 607519, 4 pages http://dx.doi.org/10.1155/2015/607519 Case Report Mixed Cutaneous Infection Caused by Mycobacterium szulgai and Mycobacterium intermedium in a Healthy Adult Female: A Rare Case Report Amresh Kumar Singh,1 Rungmei S. K. Marak,2 Anand Kumar Maurya,3 Manaswini Das,2 Vijaya Lakshmi Nag,3 and Tapan N. Dhole2 1 Department of Microbiology, BRD Medical College, Gorakhpur, Uttar Pradesh 273013, India 2Department of Microbiology, Sanjay Gandhi Post Graduate Institute of Medical Sciences, Lucknow 226014, India 3Department of Microbiology, All India Institute of Medical Sciences, Jodhpur 342005, India Correspondence should be addressed to Amresh Kumar Singh; [email protected] Received 2 December 2014; Revised 2 February 2015; Accepted 2 February 2015 Academic Editor: Kowichi Jimbow Copyright © 2015 Amresh Kumar Singh et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Nontuberculous mycobacteria (NTMs) are ubiquitous and are being increasingly reported as human opportunistic infection. Cutaneous infection caused by mixed NTM is extremely rare. We encountered the case of a 46-year-old female, who presented with multiple discharging sinuses over the lower anterior abdominal wall (over a previous appendectomy scar) for the past 2 years. Microscopy and culture of the pus discharge were done to isolate and identify the etiological agent. Finally, GenoType Mycobacterium CM/AS assay proved it to be a mixed infection caused by Mycobacterium szulgai and M.
    [Show full text]
  • The Impact of Chlorine and Chloramine on the Detection and Quantification of Legionella Pneumophila and Mycobacterium Spp
    The impact of chlorine and chloramine on the detection and quantification of Legionella pneumophila and Mycobacterium spp. Maura J. Donohue Ph.D. Office of Research and Development Center of Environmental Response and Emergency Response (CESER): Water Infrastructure Division (WID) Small Systems Webinar January 28, 2020 Disclaimer: The views expressed in this presentation are those of the author and do not necessarily reflect the views or policies of the U.S. Environmental Protection Agency. A Tale of Two Bacterium… Legionellaceae Mycobacteriaceae • Legionella (Genus) • Mycobacterium (Genus) • Gram negative bacteria • Nontuberculous Mycobacterium (NTM) (Gammaproteobacteria) • M. avium-intracellulare complex (MAC) • Flagella rod (2-20 µm) • Slow grower (3 to 10 days) • Gram positive bacteria • Majority of species will grow in free-living • Rod shape(1-10 µm) amoebae • Non-motile, spore-forming, aerobic • Aerobic, L-cysteine and iron salts are required • Rapid to Slow grower (1 week to 8 weeks) for in vitro growth, pH: 6.8 to 7, T: 25 to 43 °C • ~156 species • ~65 species • Some species capable of causing disease • Pathogenic or potentially pathogenic for human 3 NTM from Environmental Microorganism to Opportunistic Opponent Genus 156 Species Disease NTM =Nontuberculous Mycobacteria MAC = M. avium Complex Mycobacterium Mycobacterium duvalii Mycobacterium litorale Mycobacterium pulveris Clinically Relevant Species Mycobacterium abscessus Mycobacterium elephantis Mycobacterium llatzerense. Mycobacterium pyrenivorans, Mycobacterium africanum Mycobacterium europaeum Mycobacterium madagascariense Mycobacterium rhodesiae Mycobacterium agri Mycobacterium fallax Mycobacterium mageritense, Mycobacterium riyadhense Mycobacterium aichiense Mycobacterium farcinogenes Mycobacterium malmoense Mycobacterium rufum M. avium, M. intracellulare, Mycobacterium algericum Mycobacterium flavescens Mycobacterium mantenii Mycobacterium rutilum Mycobacterium alsense Mycobacterium florentinum. Mycobacterium marinum Mycobacterium salmoniphilum ( M. fortuitum, M.
    [Show full text]
  • Association of Non-Tuberculous Mycobacteria with Mycobacterium Leprae in Environment of Leprosy Endemic Regions in India T ⁎ Ravindra P
    Infection, Genetics and Evolution 72 (2019) 191–198 Contents lists available at ScienceDirect Infection, Genetics and Evolution journal homepage: www.elsevier.com/locate/meegid Research Paper Association of non-tuberculous mycobacteria with Mycobacterium leprae in environment of leprosy endemic regions in India T ⁎ Ravindra P. Turankara, , Vikram Singha, Hariom Guptaa, Vinay Kumar Pathaka, Madhvi Ahujaa, Itu Singha, Mallika Lavaniaa, Amit K. Dindab, Utpal Senguptaa a The Leprosy Mission Trust India, Stanley Browne Laboratory, TLM community Hospital Nand Nagari, New Delhi 110093, India b Department of Pathology, Institute/University: All India Institute of Medical Sciences, Ansari Nagar, New Delhi 110 029, India ARTICLE INFO ABSTRACT Keywords: Non-tuberculous mycobacteria (NTM) are environmental mycobacteria found ubiquitously in nature. The pre- Mycobacterial species sent study was conducted to find out the presence of various species of NTM in leprosy endemic region along 16S rRNA gene target with Mycobacterium (M) leprae. Water and wet soil samples from the periphery of ponds used by the community Sequencing were collected from districts of Purulia of West Bengal and Champa of Chhattisgarh, India. Samples were pro- FITC cessed and decontaminated followed by culturing on Lowenstein Jensen (LJ) media. Polymerase chain reaction PGL-1 (PCR) was performed using 16S rRNA gene target of mycobacteria and species was confirmed by sequencing M. leprae method. Indirect immune-fluorescent staining of M. leprae from soil was performed using M. leprae-PGL-1 rabbit polyclonal antibody. The phylogenetic tree was constructed by using MEGA-X software. From 380 soil samples 86 NTM were isolated, out of which 34(40%) isolates were rapid growing mycobacteria (RGM) and 52(60%) isolates were slow growing mycobacteria (SGM).
    [Show full text]
  • Nunescosta2016.Pdf
    Tuberculosis 96 (2016) 107e119 Contents lists available at ScienceDirect Tuberculosis journal homepage: http://intl.elsevierhealth.com/journals/tube REVIEW The looming tide of nontuberculous mycobacterial infections in Portugal and Brazil Daniela Nunes-Costa a, Susana Alarico a, Margareth Pretti Dalcolmo b, * Margarida Correia-Neves c, d, Nuno Empadinhas a, e, a CNC e Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal b Reference Center Helio Fraga, Fundaçao~ Oswaldo Cruz, FIOCRUZ, MoH, Rio de Janeiro, Brazil c ICVS e Health and Life Sciences Research Institute, University of Minho, Braga, Portugal d ICVS/3B's, PT Government Associate Laboratory, Braga/Guimaraes,~ Portugal e IIIUC e Institute for Interdisciplinary Research, University of Coimbra, Coimbra, Portugal article info summary Article history: Nontuberculous mycobacteria (NTM) are widely disseminated in the environment and an emerging Received 5 May 2015 cause of infectious diseases worldwide. Their remarkable natural resistance to disinfectants and anti- Received in revised form biotics and an ability to survive under low-nutrient conditions allows NTM to colonize and persist in 27 August 2015 man-made environments such as household and hospital water distribution systems. This overlap be- Accepted 16 September 2015 tween human and NTM environments afforded new opportunities for human exposure, and for expression of their often neglected and underestimated pathogenic potential. Some risk factors pre- Keywords: disposing to NTM disease have been
    [Show full text]
  • Shared Pathogenomic Patterns Characterize
    Shared Pathogenomic Patterns Characterize a New Phylotype, Revealing Transition toward Host-Adaptation Long before Speciation of Mycobacterium tuberculosis Guillaume Sapriel, Roland Brosch To cite this version: Guillaume Sapriel, Roland Brosch. Shared Pathogenomic Patterns Characterize a New Phylotype, Revealing Transition toward Host-Adaptation Long before Speciation of Mycobacterium tuberculosis. Genome Biology and Evolution, Society for Molecular Biology and Evolution, 2019, 11 (8), pp.2420- 2438. 10.1093/gbe/evz162. hal-02379155 HAL Id: hal-02379155 https://hal.archives-ouvertes.fr/hal-02379155 Submitted on 27 Jan 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Distributed under a Creative Commons Attribution| 4.0 International License GBE Shared Pathogenomic Patterns Characterize a New Phylotype, Revealing Transition toward Host-Adaptation Long before Speciation of Mycobacterium tuberculosis Guillaume Sapriel1,2,* and Roland Brosch 3 Downloaded from https://academic.oup.com/gbe/article-abstract/11/8/2420/5542391 by Institut Pasteur user on 27 January 2020 1UFR des Sciences de La Sante, Universite de Versailles St. Quentin, Montigny le Bretonneux, France 2Atelier de Bioinformatique, ISYEB, UMR 7205, Paris, France 3Unit for Integrated Mycobacterial Pathogenomics, Institut Pasteur, CNRS UMR 3525, Paris, France *Corresponding author: E-mail: [email protected].
    [Show full text]
  • Diagnosis, Treatment, and Prevention of Nontuberculous Mycobacterial Diseases
    American Thoracic Society Documents An Official ATS/IDSA Statement: Diagnosis, Treatment, and Prevention of Nontuberculous Mycobacterial Diseases David E. Griffith, Timothy Aksamit, Barbara A. Brown-Elliott, Antonino Catanzaro, Charles Daley, Fred Gordin, Steven M. Holland, Robert Horsburgh, Gwen Huitt, Michael F. Iademarco, Michael Iseman, Kenneth Olivier, Stephen Ruoss, C. Fordham von Reyn, Richard J. Wallace, Jr., and Kevin Winthrop, on behalf of the ATS Mycobacterial Diseases Subcommittee This Official Statement of the American Thoracic Society (ATS) and the Infectious Diseases Society of America (IDSA) was adopted by the ATS Board Of Directors, September 2006, and by the IDSA Board of Directors, January 2007 CONTENTS Health Care– and Hygiene-associated Disease and Disease Prevention Summary NTM Species: Clinical Aspects and Treatment Guidelines Diagnostic Criteria of Nontuberculous Mycobacterial M. avium Complex (MAC) Lung Disease Key Laboratory Features of NTM M. kansasii Health Care- and Hygiene-associated M. abscessus Disease Prevention M. chelonae Prophylaxis and Treatment of NTM Disease M. fortuitum Introduction M. genavense Methods M. gordonae Taxonomy M. haemophilum Epidemiology M. immunogenum Pathogenesis M. malmoense Host Defense and Immune Defects M. marinum Pulmonary Disease M. mucogenicum Body Morphotype M. nonchromogenicum Tumor Necrosis Factor Inhibition M. scrofulaceum Laboratory Procedures M. simiae Collection, Digestion, Decontamination, and Staining M. smegmatis of Specimens M. szulgai Respiratory Specimens M. terrae
    [Show full text]