National Reference Center for Mycobacteria

Borstel-Hohenraden, Germany

National Reference Center for Mycobacteria

Borstel-Hohenraden, Germany
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Diel R.,Hannover Medical School | Loddenkemper R.,German Central Committee Against Tuberculosis | Niemann S.,National Reference Center for Mycobacteria | Nienhaus A.,Institution for Statutory Accident Insurance and Prevention in the Health and Welfare Services
American Journal of Respiratory and Critical Care Medicine | Year: 2011

Rationale: Only limited data are available on the predictive value of interferon-γ release assays for progression from latent tuberculosis infection to active tuberculosis (TB). Objectives: To build on our initial study comparing the QuantiFERON-TB Gold in-tube assay (QFT) with the tuberculin skin test (TST) in close contacts of patients with TB and evaluating progression to active TB for up to 4 years. Methods: A cohort of close contacts of smear-positive index cases established between May 2005 and April 2008 was tested with QFT and TST. Through April 2010, progressors to active TB were consecutively recorded. Measurements and Main Results: Of the 1,414 contacts (141 children), 1,033 were still resident in Hamburg at the end of the study period, and results of both tests were available for 954. QFT, but not TST, results were associated with exposure time (P < 0.0001). For QFT, 198 of 954 (20.8%) were positive; 63.3% (604) were TST positive at greater than 5 mm and 25.4% at greater than 10 mm. Nine hundred and three contacts refused chemoprevention and 19 developed active TB. All 19 (100%) had been QFT positive with a progression rate of 12.9% (19 of 147) over the observation period. Corresponding values for the TST were significantly lower: 89.5% (17 of 19) and 3.1% (17 of 555) at greater than 5 mm, and 52.6% (10 of 19) and 4.8% (10 of 207) at greater than 10 mm, respectively. The progression rate of 28.6% (6 of 21) for QFT-positive children was significantly higher than 10.3% (13 of 126) for adults (P = 0.03). Conclusions: Results suggest that QFT is more reliable than the TST for identifying those who will soon progress to active TB, especially in children.


Roetzer A.,Molecular Mycobacteriology | Diel R.,University of Kiel | Kohl T.A.,Molecular Mycobacteriology | Kohl T.A.,Bielefeld University | And 10 more authors.
PLoS Medicine | Year: 2013

Background: Understanding Mycobacterium tuberculosis (Mtb) transmission is essential to guide efficient tuberculosis control strategies. Traditional strain typing lacks sufficient discriminatory power to resolve large outbreaks. Here, we tested the potential of using next generation genome sequencing for identification of outbreak-related transmission chains. Methods and Findings: During long-term (1997 to 2010) prospective population-based molecular epidemiological surveillance comprising a total of 2,301 patients, we identified a large outbreak caused by an Mtb strain of the Haarlem lineage. The main performance outcome measure of whole genome sequencing (WGS) analyses was the degree of correlation of the WGS analyses with contact tracing data and the spatio-temporal distribution of the outbreak cases. WGS analyses of the 86 isolates revealed 85 single nucleotide polymorphisms (SNPs), subdividing the outbreak into seven genome clusters (two to 24 isolates each), plus 36 unique SNP profiles. WGS results showed that the first outbreak isolates detected in 1997 were falsely clustered by classical genotyping. In 1998, one clone (termed "Hamburg clone") started expanding, apparently independently from differences in the social environment of early cases. Genome-based clustering patterns were in better accordance with contact tracing data and the geographical distribution of the cases than clustering patterns based on classical genotyping. A maximum of three SNPs were identified in eight confirmed human-to-human transmission chains, involving 31 patients. We estimated the Mtb genome evolutionary rate at 0.4 mutations per genome per year. This rate suggests that Mtb grows in its natural host with a doubling time of approximately 22 h (400 generations per year). Based on the genome variation discovered, emergence of the Hamburg clone was dated back to a period between 1993 and 1997, hence shortly before the discovery of the outbreak through epidemiological surveillance. Conclusions: Our findings suggest that WGS is superior to conventional genotyping for Mtb pathogen tracing and investigating micro-epidemics. WGS provides a measure of Mtb genome evolution over time in its natural host context. Please see later in the article for the Editors' Summary. © 2013 Roetzer et al.


Beckert P.,Research Center Borstel | Hillemann D.,National Reference Center for Mycobacteria | Kohl T.A.,Bielefeld University | Kalinowski J.,Bielefeld University | And 3 more authors.
Antimicrobial Agents and Chemotherapy | Year: 2012

The ribosomal L3 protein was identified as a novel target in linezolid (LZD)-resistant Mycobacterium tuberculosis strains. Next-generation sequencing confirmed rplC T460C as the sole mutation in an LZD-resistantM. tuberculosis H37Rv strain selected in vitro. Sequencing analysis revealed the rplC T460C mutation in eight further LZD-resistant isolates (three in vitro-selected mutants and five patient isolates, including isolates from three different patients that developed LZD resistance during treatment) but in none of the susceptible control strains (n = 84). Copyright © 2012, American Society for Microbiology. All Rights Reserved.


Diel R.,University of Kiel | Nienhaus A.,University of Hamburg | Lampenius N.,University of Hohenheim | Rusch-Gerdes S.,National Reference Center for Mycobacteria | Richter E.,National Reference Center for Mycobacteria
Respiratory Medicine | Year: 2014

SummaryObjectives 4220 new cases of tuberculosis (TB) were reported in Germany in 2012; of those, 65 cases were multidrug-resistant TB (MDR-TB) or extensively multidrug-resistant TB (XDR-TB) cases. However, there is only limited information on the economic consequences of drug resistance patterns on the treatment costs of MDR-and XDR-TB patients. Methods On the basis of drug susceptibility of the single MDR-TB/XDR-TB strains the direct medical costs of suitable therapies were calculated according to the current guidelines of the World Health Organization (WHO) and those of the German Central Committee against Tuberculosis. These costs were combined with hospital and outpatients monitoring costs and followed the most recent German invoicing system and health statistics. Total drug and monitoring costs and were determined by Monte-Carlo simulation comprising all different options. Results According to this, the mean drug costs were €51,113.22 (range €19,586.14 to €94,767.90). The weighted costs for hospitalization were €26,000.76 per patient compared to only €2,192.13 for primary outpatients; the total treatment costs of MDR-TB amounted to €64,429.23. These are joined by the costs due to loss of productivity, varying between €17,721.60 and €44,304. From a societal perspective, the total cost per MDR-TB/XDR-TB case reach an amount between €82,150 and €108,733 per case, respectively. Conclusion Cost analyses based on strain resistance patterns allow more reliable estimates of the real costs of treating MDR-TB/XDR-TB than do methods that ignore this factor. Advantageously, they demonstrate the economic impact of drug-resistant TB in low-incidence countries. Costs of productivity loss is of new importance because of the length of MDR-XDR therapy, but its true share of total costs has still to be determined. © 2014 The Authors. Published by Elsevier Ltd.


Feuerriegel S.,Research Center Borstel | Oberhauser B.,German Leprosy and TB Relief Association | George A.G.,National Leprosy TB Reference Laboratory | Dafae F.,Ministry of Health and Sanitation | And 3 more authors.
BMC Microbiology | Year: 2012

Background: Drug resistance displays a problem for the therapy of Mycobacterium tuberculosis infections. For molecular resistance testing, it is essential to have precise knowledge on genomic variations involved in resistance development. However, data from high-incidence settings are only sparely available. Therefore we performed a systematic approach and analyzed a total of 97 M. tuberculosis strains from previously treated patients in Sierra Leone for mutations in katG, rpoB, rrs, rpsL, gidB, embB, pncA and where applicable in inhA and ahpC. Of the strains investigated 50 were either mono- or poly-resistant to isoniazid, rifampin, streptomycin, ethambutol and pyrazinamide or MDR and 47 fully susceptible strains served as controls. Results: The majority of isoniazid and rifampin resistant strains had mutations in katG315 (71.9%) and rpoB531 (50%). However, rpoB mutations in codons 511, 516 and 533 were also detected in five rifampin susceptible strains. MIC determinations revealed low-level rifampin resistance for those strains. Thus, the sensitivity and specificity of sequencing of katG for detection of drug resistance were 86.7% and 100% and for sequencing of rpoB 100% and 93.8%, respectively. Strikingly, none of the streptomycin resistant strains had mutations in rrs, but 47.5% harboured mutations in rpsL. Further changes were detected in gidB. Among ethambutol resistant strains 46.7% had mutations at embB306. Pyrazinamide resistant strains displayed a variety of mutations throughout pncA. The specificities of sequencing of rpsL, embB and pncA for resistance detection were high (96-100%), whereas sensitivities were lower (48.8%, 73.3%, 70%). Conclusions: Our study reveals a good correlation between data from molecular and phenotypic resistance testing in this high-incidence setting. However, the fact that particular mutations in rpoB are not linked to high-level resistance is challenging and demonstrates that careful interpretation of molecular resistance assays is mandatory. In addition, certain variations, especially in gidB, appear to be phylogenetically informative polymorphisms rather than markers for drug resistance. © 2012 Feuerriegel et al.; licensee BioMed Central Ltd.


Niemann S.,National Reference Center for Mycobacteria | Diel R.,Hannover Medical School | Khechinashvili G.,Georgian Foundation Against Tuberculosis and Lung Diseases | Gegia M.,Georgian Foundation Against Tuberculosis and Lung Diseases | And 2 more authors.
Journal of Clinical Microbiology | Year: 2010

High rates and transmission of multidrug-resistant (MDR) tuberculosis (TB) have been associated with the Mycobacterium tuberculosis complex (MTBC) Beijing lineage, pointing to the importance of pathogen genetic factors for the modulation of infection outcome and epidemiology. We present here an in-depth analysis of the population structure of MTBC strains from the Republic of Georgia, a high-incidence setting at the Black Sea Coast. Phylogenetic lineages were identified based on 24-locus MIRU-VNTR (for mycobacterial interspersed repetitive unit-variable number tandem repeat) and spoligotyping analysis. Clusters of strains with identical genotyping profiles were determined as an indicator for the rate of recent transmission. Among the 183 M. tuberculosis isolates investigated, the most prominent lineage found was Beijing (26%), followed by the LAM (18%), Ural (12%), and Haarlem (5%) strains. A closely related previously undefined phylogenetic group (62 strains) showed a genotyping pattern similar to laboratory strain H37RV and was denominated as "Georgia-H37RV-like." Although isoniazid resistance was found among strains of different lineages, MDR TB was nearly completely restricted to Beijing strains (P < 0.0001). Approximately 50% of the isolates were grouped in clusters, indicating a high rate of recent transmission. Our data indicate that, in addition to the confirmation of the importance of Beijing genotype strains for the TB epidemiology in former Soviet Union countries, a high-population diversity with strains of the LAM, Ural, Haarlem, and a previously undefined lineage represents nearly two-thirds of the strains found in Georgia. Higher rates among previously treated and MDR TB patients point to a higher potential of lineage Beijing to escape therapy and develop MDR TB. Copyright © 2010, American Society for Microbiology. All Rights Reserved.


Prodinger W.M.,Innsbruck Medical University | Indra A.,AGES Institute for Medical Microbiology and Hygiene Vienna | Koksalan O.K.,Istanbul University | Kilicaslan Z.,Istanbul University | Richter E.,National Reference Center for Mycobacteria
Expert Review of Anti-Infective Therapy | Year: 2014

Mycobacterium caprae, a member of the Mycobacterium tuberculosis complex, causes tuberculosis (TB) in man and animals. Some features distinguish M. caprae from its epidemiological twin, Mycobacterium bovis: M. caprae is evolutionarily older, accounts for a smaller burden of zoonotic TB and is not globally distributed, but primarily restricted to European countries. M. caprae occurs only in a low proportion of human TB cases and this proportion may even decrease, if progress toward eradication of animal TB in Europe continues. So why bother, if M. caprae is not an enigma for diagnostic TB tests and if resistance against first-line drugs is a rarity with M. caprae? This 'European' pathogen of zoonotic TB asks interesting questions regarding the definition of a species. The latter, seemingly only an academic question, particularly requires and challenges the collaboration between human and veterinary medicine. © 2014 Informa UK Ltd.


Genc G.E.,Istanbul University | Richter E.,National Reference Center for Mycobacteria | Erturan Z.,Istanbul University
APMIS | Year: 2013

Nontuberculous mycobacteria (NTM) are ubiquitous in hot and cold water distribution systems. With molecular typing methods it was shown that water can be the source of colonization and infection with NTM. The aim of our study was the investigation of NTM in hot and cold water samples taken from various departments of two hospitals in Istanbul, Turkey. Totally, 160 water samples were examined. The temperature, pH, and free chlorine levels of water samples were measured between 10-41 °C, 6.78-7.98 and <0.3-0.5 mg/L, respectively. NTM were detected in 33 (20.6%) samples. Totally 20 (60.6%), 10 (30.3%) and 3 (9.1%) isolates were identified as Mycobacterium lentiflavum, Mycobacterium gordonae, and Mycobacterium peregrinum, respectively. M. lentiflavum, which was the most frequently isolated NTM, is characterized by multiple resistance to antimycobacterial drugs. Although no infections with this mycobacterium were reported from our country so far, preventive measures may be considered in patients under immunosuppression. Because no significant correlations were found among the presence of NTM or species distribution and water temperature, pH or free chlorine levels, other factors need to be investigated. © 2013 APMIS.


Napiorkowska A.,National Tuberculosis and Lung Diseases Research Institute | Rusch-Gerdes S.,National Reference Center for Mycobacteria | Hillemann D.,National Reference Center for Mycobacteria | Richter E.,National Reference Center for Mycobacteria | Augustynowicz-Kopec E.,National Tuberculosis and Lung Diseases Research Institute
International Journal of Tuberculosis and Lung Disease | Year: 2014

BACKGROUND: Pyrazinamide (PZA) is an important first-line anti-tuberculosis drug that is generally administered with isoniazid, rifampicin, ethambutol and streptomycin. OBJECTIVE: To analyse the correlation between phenotypic resistance to PZA and genotype to find out whether the great diversity in pncA mutations is epidemiologically useful in tracing the transmission of PZA-resistant Mycobacterium tuberculosis strains among patients. MATERIALS AND METHODS: The study included 71 PZA-resistant M. tuberculosis strains isolated from 62 Polish and 9 German patients. All strains were analysed using minimal inhibitory concentration value determination, pncA mutation analysis, spoligotyping, 24-loci mycobacterial interspersed repetitive units-variable number of tandem repeats (MIRU-VNTR) and insertion sequence (IS) 6110 restriction fragment length polymorphism (RFLP) fingerprinting. RESULTS: In 63 isolates, 37 (88.7%) different mutations in the pncA gene were observed, 13 of which had not been previously reported; 11 molecular families with the same MIRU-VNTR and IS6110-RFLP pattern were found. The same mutation was identified in three families, while different ones were identified in the remaining families. CONCLUSION: Mutations in the pncA gene are a major cause of PZA resistance in M. tuberculosis. pncA mutation analysis can be used to obtain valuable additional information, but should be applied with caution for epidemiological analysis. © 2014 The Union.


Andres S.,National Reference Center for Mycobacteria | Hillemann D.,National Reference Center for Mycobacteria | Rusch-Gerdes S.,National Reference Center for Mycobacteria | Richter E.,National Reference Center for Mycobacteria
Antimicrobial Agents and Chemotherapy | Year: 2014

Four out of 143 phenotypically isoniazid-resistant but rifampin-susceptible Mycobacterium tuberculosis strains that were isolated from patients in Germany in 2011 had mutations in the rifampin resistance-determining region of rpoB. After performing drug susceptibility testing (DST) with two methods, the proportion method on Löwenstein-Jensen medium and using the Bactec 960 Mycobacteria Growth Indicator Tube system, we conclude that the two methods are equally reliable for phenotypic DST and MIC determination. © 2014, American Society for Microbiology. All Rights Reserved.

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