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  • American Society for Microbiology  (11)
  • 1
    Online Resource
    Online Resource
    Some Synonymous and Nonsynonymous gyrA Mutations in Mycobacterium tuberculosis Lead to Systematic False-Positive Fluoroquinolone Resistance Results with the Hain GenoType MTBDR sl Assays
    American Society for Microbiology ; 2017
    In:  Antimicrobial Agents and Chemotherapy Vol. 61, No. 4 ( 2017-04)
    Details
    In: Antimicrobial Agents and Chemotherapy, American Society for Microbiology, Vol. 61, No. 4 ( 2017-04)
    Abstract: In this study, using the Hain GenoType MTBDR sl assays (versions 1 and 2), we found that some nonsynonymous and synonymous mutations in gyrA in Mycobacterium tuberculosis result in systematic false-resistance results to fluoroquinolones by preventing the binding of wild-type probes. Moreover, such mutations can prevent the binding of mutant probes designed for the identification of specific resistance mutations. Although these mutations are likely rare globally, they occur in approximately 7% of multidrug-resistant tuberculosis strains in some settings.
    Type of Medium: Online Resource
    ISSN: 0066-4804 , 1098-6596
    URL: Article
    DOI: 10.1128/AAC.02169-16
    RVK:
    XA 24552
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 2017
    detail.hit.zdb_id: 1496156-8
    SSG: 12
    SSG: 15,3
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  • 2
    Online Resource
    Online Resource
    Role of Alanine Racemase Mutations in Mycobacterium tuberculosis d -Cycloserine Resistance
    American Society for Microbiology ; 2017
    In:  Antimicrobial Agents and Chemotherapy Vol. 61, No. 12 ( 2017-12)
    Details
    In: Antimicrobial Agents and Chemotherapy, American Society for Microbiology, Vol. 61, No. 12 ( 2017-12)
    Abstract: A screening of more than 1,500 drug-resistant strains of Mycobacterium tuberculosis revealed evolutionary patterns characteristic of positive selection for three alanine racemase (Alr) mutations. We investigated these mutations using molecular modeling, in vitro MIC testing, as well as direct measurements of enzymatic activity, which demonstrated that these mutations likely confer resistance to d -cycloserine.
    Type of Medium: Online Resource
    ISSN: 0066-4804 , 1098-6596
    URL: Article
    DOI: 10.1128/AAC.01575-17
    RVK:
    XA 24552
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 2017
    detail.hit.zdb_id: 1496156-8
    SSG: 12
    SSG: 15,3
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  • 3
    Online Resource
    Online Resource
    Epidemiology of Tuberculosis in Hamburg, Germany: Long-Term Population-Based Analysis Applying Classical and Molecular Epidemiological Techniques
    American Society for Microbiology ; 2002
    In:  Journal of Clinical Microbiology Vol. 40, No. 2 ( 2002-02), p. 532-539
    Details
    In: Journal of Clinical Microbiology, American Society for Microbiology, Vol. 40, No. 2 ( 2002-02), p. 532-539
    Abstract: To determine a detailed picture of tuberculosis (TB) epidemiology in Hamburg, Germany, 423 Mycobacterium tuberculosis complex isolates from 77.0% of all patients with culture-confirmed TB diagnosed from 1997 to 1999 in Hamburg were analyzed by IS 6110 DNA fingerprinting. IS 6110 restriction fragment length polymorphism (RFLP) clusters were assumed to have arisen from recent transmission. Results of contact tracing and additional patient interviews were used for further epidemiological analyses. Of the 423 cases, 398 were included in the cluster analysis, of which 135 (33.9%) were classified into 35 clusters ranging from 2 to 23 patients. Epidemiological links verifying recent transmission could be confirmed for 87 of the 135 clustered patients. Risk factors for recent transmission were calculated by a two-step procedure: first, based on patients with clustered isolates; and second, based on patients with clustered isolates and transmission links. In both analyses, alcohol abuse appeared to be the strongest predictor for recent transmission, followed by a history of previous contact tracing and unemployment. Homelessness, foreign ethnicity, sex, drug addiction, and human immunodeficiency virus positivity were not independent risk factors for clustering in multivariate analyses. Classical contact tracing performed prior to IS 6110 RFLP analysis identified only 24 of the 135 clustered patient. In conclusion, recent transmission seems to be frequent in Hamburg and was found to be strongly associated with alcohol abuse. Conventional contact tracing appears to be insufficient for the detection of recent transmission chains. The data presented also indicate that improved TB control strategies, including the use of RFLP for the detection of transmission chains, are needed for TB control in the setting of countries with a low incidence of TB.
    Type of Medium: Online Resource
    ISSN: 0095-1137 , 1098-660X
    URL: Article
    DOI: 10.1128/JCM.40.2.532-539.2002
    RVK:
    XA 10000
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 2002
    detail.hit.zdb_id: 1498353-9
    SSG: 12
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  • 4
    Online Resource
    Online Resource
    Assessment of an Optimized Mycobacterial Interspersed Repetitive- Unit-Variable-Number Tandem-Repeat Typing System Combined with Spoligotyping for Population-Based Molecular Epidemiology Studies of Tuberculosis
    American Society for Microbiology ; 2007
    In:  Journal of Clinical Microbiology Vol. 45, No. 3 ( 2007-03), p. 691-697
    Details
    In: Journal of Clinical Microbiology, American Society for Microbiology, Vol. 45, No. 3 ( 2007-03), p. 691-697
    Abstract: An optimized set of 24 mycobacterial interspersed repetitive-unit-variable-number tandem-repeat (MIRU-VNTR) loci, including a discriminatory subset of 15 loci, has recently been defined for the typing of Mycobacterium tuberculosis . Here, we evaluated the performances of this MIRU-VNTR typing system in combination with spoligotyping for the detection of transmission chains in a population-based study comprising 91% of culture-confirmed tuberculosis patients reported in 2003 in Hamburg, Germany. Of the 154 isolates investigated, more than 90% had high IS 6110 copy numbers (≥6). IS 6110 restriction fragment length polymorphism (RFLP) typing resulted in 13 clusters, 5 of which had a confirmed epidemiological link. All five, as well as six of the eight IS6110 clusters with no identified epidemiological link, were perfectly matched by MIRU-VNTR typing with the 24 loci. Two IS 6110 clusters were split by differences into 6 to 12 MIRU-VNTR loci, clearly supporting the absence of a link, as judged by contact tracing data. In contrast, only one MIRU-VNTR cluster, grouping what were probably epidemiologically unlinked isolates, was split by IS 6110 RFLP. However, these isolates were also distinguished by spoligotyping. Both the optimized 24-locus and 15-locus sets thus showed a comparable to slightly better predictive value, especially when combined with spoligotyping, than the current gold standard IS 6110 RFLP for the study of tuberculosis transmission in Hamburg. Because the epidemiological characteristics of this setting are similar to those of many developed countries, these results support the wide applicability of this real-time genotyping approach for population-based studies of M. tuberculosis transmission.
    Type of Medium: Online Resource
    ISSN: 0095-1137 , 1098-660X
    URL: Article
    DOI: 10.1128/JCM.01393-06
    RVK:
    XA 10000
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 2007
    detail.hit.zdb_id: 1498353-9
    SSG: 12
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  • 5
    Online Resource
    Online Resource
    Mycobacterium tuberculosis Beijing Lineage Favors the Spread of Multidrug-Resistant Tuberculosis in the Republic of Georgia
    American Society for Microbiology ; 2010
    In:  Journal of Clinical Microbiology Vol. 48, No. 10 ( 2010-10), p. 3544-3550
    Details
    In: Journal of Clinical Microbiology, American Society for Microbiology, Vol. 48, No. 10 ( 2010-10), p. 3544-3550
    Abstract: 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.
    Type of Medium: Online Resource
    ISSN: 0095-1137 , 1098-660X
    URL: Article
    DOI: 10.1128/JCM.00715-10
    RVK:
    XA 10000
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 2010
    detail.hit.zdb_id: 1498353-9
    SSG: 12
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  • 6
    Online Resource
    Online Resource
    Evaluation of Mycobacterium tuberculosis Typing Methods in a 4-Year Study in Schleswig-Holstein, Northern Germany
    American Society for Microbiology ; 2011
    In:  Journal of Clinical Microbiology Vol. 49, No. 12 ( 2011-12), p. 4173-4178
    Details
    In: Journal of Clinical Microbiology, American Society for Microbiology, Vol. 49, No. 12 ( 2011-12), p. 4173-4178
    Abstract: In order to evaluate the discriminatory power of different methods for genotyping of Mycobacterium tuberculosis complex (MTBC) isolates, we compared the performance of (i) IS 6110 DNA fingerprint typing, (ii) spoligotyping, and (iii) 24-loci mycobacterial interspersed repetitive units-variable number of tandem repeats (MIRU-VNTR) typing in a long-term study on the epidemiology of tuberculosis (TB) in Schleswig-Holstein, the northernmost federal state of Germany. In total, we analyzed 277 MTBC isolates collected from patients between the years 2006 and 2010. The collection comprised a broad spectrum of 13 different genotypes, among which strains of the Haarlem genotype (31%) were most prominent, followed by strains belonging to the Delhi and Beijing lineages (7% and 6%, respectively). On the basis of IS 6110 restriction fragment length polymorphism (RFLP) and spoligotyping analyses, 211 isolates had unique patterns (76%) and 66 isolates (24%) were in 20 clusters. MIRU-VNTR combined with spoligotyping analyses revealed 202 isolates with unique patterns (73%) and 75 isolates in 18 clusters (27%). Overall, there was 93.1% concordance between the typing results obtained; 198 strains were identified as unique, and 60 isolates were clustered by both typing combinations (including all 31 isolates with confirmed epidemiological links). Of the remaining 19 isolates with discrepant results, 15 were falsely clustered by MIRU-VNTR (six Beijing genotype strains) and four were clustered by IS 6110 RFLP (low IS 6110 copy number) only. In conclusion, in the study population investigated, a minority of isolates, especially of the Beijing genotype, clustered by standard 24-loci MIRU-VNTR and without an obvious epidemiological link may require second-line typing by IS 6110 RFLP or hypervariable MIRU-VNTR loci.
    Type of Medium: Online Resource
    ISSN: 0095-1137 , 1098-660X
    URL: Article
    DOI: 10.1128/JCM.05293-11
    RVK:
    XA 10000
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 2011
    detail.hit.zdb_id: 1498353-9
    SSG: 12
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  • 7
    Online Resource
    Online Resource
    Molecular Epidemiology of Tuberculosis among Immigrants in Hamburg, Germany
    American Society for Microbiology ; 2004
    In:  Journal of Clinical Microbiology Vol. 42, No. 7 ( 2004-07), p. 2952-2960
    Details
    In: Journal of Clinical Microbiology, American Society for Microbiology, Vol. 42, No. 7 ( 2004-07), p. 2952-2960
    Abstract: To study the characteristics of tuberculosis (TB) in foreign-born individuals living in Hamburg, Germany, and to discover in what way foreign-born individuals contribute to the incidence of TB in Hamburg, an in-depth epidemiological study was performed by a combination of classical and molecular methods. In total, 796 patients with TB were analyzed between 1997 and 2002 (88.7% of all patients with culture-positive TB in the study period). Of this total, 334 were foreign-born patients from 43 different countries. Of these, only 31 cases were identified as a consequence of the screening of 12,176 asylum seekers at entry. Of the foreign-born patients, 41.9% had been living in Germany for more than 5 years. On the basis of the IS 6110 typing results for isolates from all patients, 246 patients (31%) were classified into 68 clusters, with each cluster containing from 2 to 38 patients. Among foreign-born individuals, 86 (26%) were represented in 40 clusters. In multivariate analyses, a previous history as a TB contact had the highest predictive risk for clustering among foreign-born patients, followed by drug addiction, alcohol dependence, being an asylum seeker, and unemployment. Epidemiological links verifying recent transmission could be confirmed for 39 of the 86 foreign-born members (45.3%) who formed a cluster, comprising 16 source patients and 23 directly infected patients. Of 2,227 previously known contacts of foreign-born patients subjected to traditional contact investigation, 14 foreign-born individuals (0.6%) subsequently contracted culture-confirmed TB, but only 9 transmissions could be confirmed by IS 6110 typing (39.1% of the 23 confirmed fresh infections retrospectively confirmed by IS 6110 typing). In conclusion, only a minority of TB cases among foreign-born individuals are detected by screening of asylum seekers or conventional contact tracing. Recent transmission does not play an important role in TB among immigrants in Hamburg.
    Type of Medium: Online Resource
    ISSN: 0095-1137 , 1098-660X
    URL: Article
    DOI: 10.1128/JCM.42.7.2952-2960.2004
    RVK:
    XA 10000
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 2004
    detail.hit.zdb_id: 1498353-9
    SSG: 12
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  • 8
    Online Resource
    Online Resource
    Clade-Specific Virulence Patterns of Mycobacterium tuberculosis Complex Strains in Human Primary Macrophages and Aerogenically Infected Mice
    American Society for Microbiology ; 2013
    In:  mBio Vol. 4, No. 4 ( 2013-08-30)
    Details
    In: mBio, American Society for Microbiology, Vol. 4, No. 4 ( 2013-08-30)
    Abstract: Clinical strains of the Mycobacterium tuberculosis complex (MTBC) are genetically more diverse than previously anticipated. Our analysis of mycobacterial growth characteristics in primary human macrophages and aerogenically infected mice shows that the MTBC genetic differences translate into pathogenic differences in the interaction with the host. Our study reveals for the first time that “TB is not TB,” if put in plain terms. We are convinced that it is very unlikely that a single molecular mechanism may explain the observed effects. Our study refutes the hypothesis that there is a simple correlation between cytokine induction as a single functional parameter of host interaction and mycobacterial virulence. Instead, careful consideration of strain- and lineage-specific characteristics must guide our attempts to decipher what determines the pathological potential and thus the outcomes of infection with MTBC, one of the most important human pathogens.
    Type of Medium: Online Resource
    ISSN: 2161-2129 , 2150-7511
    URL: Article
    DOI: 10.1128/mBio.00250-13
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 2013
    detail.hit.zdb_id: 2557172-2
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  • 9
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    Replacing Reverse Line Blot Hybridization Spoligotyping of the Mycobacterium tuberculosis Complex
    American Society for Microbiology ; 2010
    In:  Journal of Clinical Microbiology Vol. 48, No. 5 ( 2010-05), p. 1520-1526
    Details
    In: Journal of Clinical Microbiology, American Society for Microbiology, Vol. 48, No. 5 ( 2010-05), p. 1520-1526
    Abstract: Spoligotyping is a tool for the molecular characterization/typing of Mycobacterium tuberculosis complex (MTBC) strains based on target sequences (spacers) in the direct repeat (DR) region (14). The standard spoligotyping assay involves the hybridization of amplified sample DNA to nylon membrane-immobilized oligonucleotides whose sequences are representative of 43 spacer regions. Variations in the number of spacers as a result of deletions of adjacent blocks of repetitive units allow the differentiation of clinical isolates. In the present study, we developed a new multiplexed primer extension-based spoligotyping assay using automated matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) that improves the classical reverse line blot hybridization assay with respect to reproducibility, throughput, process flow, ease of use, and data analysis. Validation of the MALDI-TOF MS-based spoligotyping assay with two sample sets with a total of 326 samples resulted in 96.6% concordance (315/326) when the full spoligotype patterns were compared with the results of standard spoligotyping and 99.9% concordance when the results were compared with those of individual primer extension assays. Ten strains (including two Mycobacterium canettii strains) showed discordant results with one or two spacer differences from the membrane-based spoligotyping result. Most discordant samples were identified to be the result of ambiguities in the interpretation of weak hybridization signals in the reverse line blot assay and sequence variations in the spacer regions. We established a new automated primer extension assay and successfully validated it for use for the routine typing of MTBC strains in the research and public health laboratory environments. The present multiplex levels of up to 30 are extendable and allow the additional incorporation of controls and antibiotic resistance markers.
    Type of Medium: Online Resource
    ISSN: 0095-1137 , 1098-660X
    URL: Article
    DOI: 10.1128/JCM.02299-09
    RVK:
    XA 10000
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 2010
    detail.hit.zdb_id: 1498353-9
    SSG: 12
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  • 10
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    Online Resource
    Stability of IS 6110 Restriction Fragment Length Polymorphism Patterns of Mycobacterium tuberculosis Strains in Actual Chains of Transmission
    American Society for Microbiology ; 2000
    In:  Journal of Clinical Microbiology Vol. 38, No. 7 ( 2000-07), p. 2563-2567
    Details
    In: Journal of Clinical Microbiology, American Society for Microbiology, Vol. 38, No. 7 ( 2000-07), p. 2563-2567
    Abstract: The stability of IS 6110 restriction fragment length polymorphism (RFLP) patterns of Mycobacterium tuberculosis strains in actual transmission chains has been assessed by analyzing the variability of IS 6110 RFLP patterns of strains in fingerprint clusters that have been confirmed by classical epidemiological data. Forty susceptible and 35 drug-resistant (including 17 multidrug-resistant) M. tuberculosis strains obtained from 75 patients living in Germany have been analyzed. The epidemiological relationship among strains within the fingerprint clusters has been verified by family contacts (14 clusters) or by contact tracing of the public health offices (7 clusters). The time spans between the first and the last isolate of one cluster ranged from less than 1 to 29 months. Of the 75 strains only 1 showed a one-band variation when compared to the other nine isolates grouped in the same cluster, corresponding with a rate of change of ∼1.9% per possible transmission (one index patient per cluster was subtracted from the total number of isolates). These results confirm a high degree of stability of IS 6110 RFLP patterns of transmitted M. tuberculosis strains. Furthermore, the data presented indicate that isolates with identical IS 6110 DNA fingerprint patterns are a good indicator for the rate of recent transmission in a study population.
    Type of Medium: Online Resource
    ISSN: 0095-1137 , 1098-660X
    URL: Article
    DOI: 10.1128/JCM.38.7.2563-2567.2000
    RVK:
    XA 10000
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 2000
    detail.hit.zdb_id: 1498353-9
    SSG: 12
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