First Insight into Genetic Diversity of the
            <i>Mycobacterium tuberculosis</i>
            Complex in Albania Obtained by Multilocus Variable-Number Tandem-Repeat Analysis and Spoligotyping Reveals the Presence of Beijing Multidrug-Resistant Isolates

Tafaj, Silva; Zhang, Jian; Hauck, Yolande; Pourcel, Christine; Hafizi, Hasan; Zoraqi, Grigor; Sola, Christophe

doi:10.1128/jcm.02284-08

Cited by 15 publications

(13 citation statements)

References 21 publications

(20 reference statements)

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“…The 24-loci MIRU-VNTR method was performed using an electrophoretic gel based method in Palermo, Italy [17]. All results were transferred into Excel spreadsheet files and imported into the Bionumerics v. 6.4 software (Applied Maths, St Martins Latem, Belgium).…”

Section: Methodsmentioning

confidence: 99%

Molecular epidemiology of tuberculosis in Sicily, Italy: what has changed after a decade?

et al. 2014

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BackgroundWe aimed to investigate the molecular epidemiology of Mycobacterium tuberculosis complex (MTBC) isolates in the province of Palermo, Sicily, Italy, by characterizing 183 isolates identified in the years 2004-2012. A comparison with 104 MTBC strains identified in the same geographic area in the years 1994-2000 was also carried out.MethodsOne hundred eighty-three MTBC isolates identified in Palermo, Italy, in the years 2004-2012 were analyzed by spoligotyping and the 24 mycobacterial interspersed repetitive unit (MIRU)-variable-number tandem-repeat (VNTR) method typing. Susceptibility testing to streptomycin, isoniazid, rifampin and ethambutol was also performed. Furthermore, the spoligotyping dataset obtained from 104 MTBC isolates identified from 1994 to 2000 was reanalyzed. Distribution into lineages and clustering of isolates in the two periods was compared.ResultsOne hundred seventy-seven out of the 183 isolates of MTBC submitted to molecular typing were fully characterized. Of these, 108 were from Italian-born and 69 from foreign-born individuals. Eleven different lineages and 35 families-subfamilies were identified with the most represented lineages being Haarlem (26.5%), T (19.2%), LAM (13.6%) and S (8.5%). Except for the Haarlem lineage, where isolates from foreign-born patients were overrepresented, the distribution of isolates in the families belonging to the Euro-American clone reflected the proportions of the two subpopulations. A total of 27 (15.2%) strains were clustered and three clusters were mixed. Approximately 25% of the 183 MTBC isolates under study proved to be resistant to at least one antiTB drug, with only three isolates categorized as multidrug resistant (MDR). When MTBC isolates identified in the years 1994-2000 were reanalyzed, lineages T (30.8%), LAM (29.8%), Haarlem (16.3%) and S (13.5%) proved to be predominant. No MTBC isolates belonging to CAM, U, CAS, Turkish and Ural lineages were identified.ConclusionsA wide heterogeneity was detected among the MTBC strains isolated in the years 2004-2012. Six lineages were not present among the isolates of the period 1994-2000. Comparison between distribution of lineages in the two consecutive periods depicts rapid and deep changes in the TB epidemiology in Palermo, Italy. An universal and continued laboratory-based surveillance of TB in Sicily is required.Electronic supplementary materialThe online version of this article (doi:10.1186/s12879-014-0602-4) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

Molecular epidemiology of tuberculosis in Sicily, Italy: what has changed after a decade?

et al. 2014

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“…These estimations were obtained keeping in mind that: (i) SIT4 cannot be assigned to any family based on spoligotyping alone; and (ii) LAM-TUR and LAM-CAM do not belong to LAM, but rather represent separate families within the Euro-American lineage. It appears that similar to Bulgaria (4%), the LAM rate is low/negligible in other Balkan countries: Turkey -3.4%; Greece -1-4%; Romania -0% (SITVIT_WEB); and Albania -0% [19]. In contrast, the LAM rate is higher in the northeastern neighbors of Bulgaria beyond the Balkans, reaching 17% in Southern Ukraine [20], 25%-39% in European Russia, and 42% in Belarus [9].…”

Section: Resultsmentioning

confidence: 67%

Prevalence of Latin-American-Mediterranean genetic family in population structure of Mycobacterium tuberculosis in Bulgaria

Valcheva¹,

Rastogi²,

Mokrousov³

2015

International Journal of Mycobacteriology

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Tuberculosis (TB) control remains an important public health priority for Bulgaria. The population structure of Mycobacterium tuberculosis is clonal and certain genetic families of this species (e.g., Latin-American-Mediterranean [LAM]) have attracted more attention due to their global dissemination and/or particular pathogenic properties, e.g., association with multidrug resistance (MDR). The aim of this study was to evaluate the prevalence of the M. tuberculosis LAM family in Bulgaria based on the use of different molecular markers. A total of 101 previously spoligotyped M. tuberculosis strains were studied by LAM-specific PCR assay to detect an insertion of IS6110 in the specific genome region. On the whole, clear-cut results were obtained for most strains; spoligotype-based family was reassigned in some of them. At the same time, double bands were amplified in some cases and warrant further validation studies of this method. The higher MDR rate among LAM versus other genotype isolates was observed (P=0.04). In conclusion, these results suggest a low (<4%) prevalence rate of LAM in Bulgaria (that is similar to its Balkan neighbors) and highlight the importance of using robust markers for correct detection of the LAM family.

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“…The mutations were already known for the other 4 strains from Germany. PCR was prepared as published previously (33) and sent to Beckman Coulter Genomics (Takeley, United Kingdom) for Sanger sequencing. Sequencing results were aligned by MultAlign (multiple sequence alignment with hierarchical clustering) (9), and the presence/absence of mutations was analyzed.…”

Section: Methodsmentioning

confidence: 99%

“…DNAs were extracted by the cetyltrimethylammonium bromide (CTAB) method or by the thermolyzate method (33,36). One hundred fourteen DNAs corresponding to 114 M. tuberculosis clinical isolates were used.…”

Section: Methodsmentioning

confidence: 99%

“Spoligoriftyping,” a Dual-Priming-Oligonucleotide-Based Direct-Hybridization Assay for Tuberculosis Control with a Multianalyte Microbead-Based Hybridization System

et al. 2012

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We developed “spoligoriftyping,” a 53-plex assay based on two preexisting methods, the spoligotyping and “rifoligotyping” assays, by combining them into a single assay. Spoligoriftyping allows simultaneous spoligotyping (i.e., clustered regularly interspaced short palindromic repeat [CRISPR]-based genotyping) and characterization of the main rifampin drug resistance mutations on the rpoB hot spot region in a few hours. This test partly uses the dual-priming-oligonucleotide (DPO) principle, which allows simultaneous efficient amplifications of rpoB and the CRISPR locus in the same sample. We tested this method on a set of 114 previously phenotypically and genotypically characterized multidrug-resistant (MDR) Mycobacterium tuberculosis or drug-susceptible M. tuberculosis DNA extracted from clinical isolates obtained from patients from Bulgaria, Nigeria, and Germany. We showed that our method is 100% concordant with rpoB sequencing results and 99.95% (3,911/3,913 spoligotype data points) correlated with classical spoligotyping results. The sensitivity and specificity of our assay were 99 and 100%, respectively, compared to those of phenotypic drug susceptibility testing. Such assays pave the way to the implementation of locally and specifically adapted methods of performing in a single tube both drug resistance mutation detection and genotyping in a few hours.

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First Insight into Genetic Diversity of the Mycobacterium tuberculosis Complex in Albania Obtained by Multilocus Variable-Number Tandem-Repeat Analysis and Spoligotyping Reveals the Presence of Beijing Multidrug-Resistant Isolates

Cited by 15 publications

References 21 publications

Molecular epidemiology of tuberculosis in Sicily, Italy: what has changed after a decade?

Molecular epidemiology of tuberculosis in Sicily, Italy: what has changed after a decade?

Prevalence of Latin-American-Mediterranean genetic family in population structure of Mycobacterium tuberculosis in Bulgaria

“Spoligoriftyping,” a Dual-Priming-Oligonucleotide-Based Direct-Hybridization Assay for Tuberculosis Control with a Multianalyte Microbead-Based Hybridization System

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