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1

Application of FLiP method for differentiation of Mycobacterium tuberculosis strains in comparison to commonly used methods, spoligotyping and MIRU-VNTR typing

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Zaczek A., Brzostek A., Gorna A., Sajduda A., Wojtasik A., Dziadek J.
Polish Journal of Microbiology
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2013
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tom 62
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nr 1
The current “gold standard” in molecular epidemiological studies of Mycobacterium tuberculosis is IS6110 RFLP based on IS6110 polymorphism. However PCR-based methods are becoming increasingly important. Recently, fast ligation-mediated PCR (FLiP), based on IS6110 polymorphism was proposed. In this study, the discriminatory power of FLiP, spoligotyping and MIRU-VNTR typing, in differentiation of M. tuberculosis isolates was compared. The discriminatory index (HGI) of spoligotyping, MIRU-VNTR analysis, and FLiP was 0.653, 0.837, and 0.917, respectively. This indicates that FLiP allows a high level of differentiation among M. tuberculosis strains and it might be a valuable alternative to the other typing methods.
2

Analysis of bovine tuberculosis transmission in Jalisco, Mexico through whole-genome sequencing

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Verdugo Escarcega D. A., Perea Razo C. A., Gonzalez Ruiz S., Sosa Gallegos S. L., Suazo F. M., Alarcon G. J. C.
Journal of Veterinary Research
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2020
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tom 64
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nr 1
3

Characterisation of Mycobacterium bovis strains isolated from farm and wild animals in Poland

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Augustynowicz-Kopec E., Krajewska M., Zabost A., Napiorkowska A., Zwolska Z.
Bulletin of the Veterinary Institute in Puławy
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2011
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tom 55
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nr 3
Sixty-one isolates of M. bovis (58 from cattle and three from wild animals) from eight regions of Poland were analysed. Molecular analysis was done using HAIN and spoligotyping methods. Drug susceptibility of the isolates to streptomycin, izoniazid, rifampicin, ethambutol, and pyrazinamide was tested by proportional methods on solid and liquid media. By spoligotyping, 47 (77%) isolates were identified as M. bovis subsp. bovis and 14 (23%) isolates were identified as M. bovis subsp. caprae. Eleven animals of the domestic cattle (18%) and all wild animals were infected by M. bovis subsp. caprae. Among cattle infected by M. bovis, 12 spoligotypes were identified, most of them not registered in the SpolDB4 database. The strains isolated from 15 animals of the domestic cattle were the same spoligo pattern. In conclusion, transmission of mycobacteria among the farm and wild animals has been suspected.
4

A two-step strategy for molecular typing of multidrug-resistant Mycobacterium tuberculosis clinical isolates from Poland

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Jagielski T., Augustynowicz-Kopec E., Pawlik K., Dziadek J., Zwolska Z., Bielecki J.
Polish Journal of Microbiology
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2011
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tom 60
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nr 3
Tuberculosis (TB) continues to be one of the most challenging public health problems in the world. An important contributor to the global burden of the disease is the emergence and spread of drug-resistant and particularly multidrug-resistant Mycobacterium tuberculosis strains (MDR), defined as being resistant to at least isoniazid and rifampicin. In recent years, the introduction of different DNA-based molecular typing methods has substantially improved the knowledge of the epidemiology of TB. The purpose of this study was to employ a combination of two PCR-based genotyping methods, namely spoligotyping and IS6110-Mtb1/Mtb2 PCR to investigate the clonal relatedness of MDR M. tuberculosis clinical isolates recovered from pulmonary TB patients from Poland. Among the 50 isolates examined, 28 (56%) were clustered by spoligotyping, whereas IS6110-Mtb1/Mtb2 PCR resulted in 16 (32%) clustered isolates. The isolates that clustered in both typing methods were assumed to be clonally related. A two-step strategy consisting of spoligotyping as a first-line test, performed on the entire pool of isolates, and IS6110-Mtb1/Mtb2 PCR typing as a confirmatory subtyping method, performed only within spoligotype-defined clusters, is an efficient approach for determining clonal relatedness among M. tuberculosis clinical isolates.
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