A Two-Step Strategy for Molecular Typing of Multidrug-Resistant Mycobacterium tuberculosis Clinical Isolates from Poland

Abstract

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.

Comparing IS6110‐RFLP, PGRS‐RFLP and IS6110‐Mtb1/Mtb2 PCR methods for genotyping ofMycobacterium tuberculosisisolates

“…In addition, other studies revealed that the calculated discriminatory power of IS6110‐Mtb1/Mtb2 PCR was greater than those of MIRU‐VNTR typing and spoligotyping and almost equivalent to that of IS6110‐RFLP genotyping (Jagielski et al . 2011, 2016). However, in a study in Poland, the differentiating power of clustering via IS6110‐Mtb1‐Mtb2 PCR was only slightly minor than RFLP‐IS6110 (Sajduda et al .…”

“…The patterns obtained from IS6110‐Mtb1/Mtb2 PCR typing using each of the combinations of primers (Mtb1‐IS1‐IS2 or Mtb2‐IS1‐IS2), were analysed separately with Lab Works 4.5 software (UVP Inc., Upland, CA) and by visual inspection as described by Jagielski et al . (2011).…”

Current Methods in the Molecular Typing ofMycobacterium tuberculosisand Other Mycobacteria

“…Although the IS 6110 -Mtb1-Mtb2 PCR method has not been exploited sufficiently, the results from the insofar performed studies suggest its usefulness in the molecular epidemiological investigations of TB. Most notably the discriminatory power of this method has been demonstrated as almost equivalent to that of IS 6110 -RFLP typing [ 67 , 148 , 149 ].…”

ANÁLISE MOLECULAR DE Mycobacterium tuberculosis ISOLADOS DE PACIENTES ATENDIDOS EM GOIÂNIA, GOIÁS, POR MEIO DO RFLP-IS6110 E DO 15 loci MIRU-VNTR

“…Os métodos de tipagem molecular vêm desempenhando, atualmente, um importante papel na capacidade de discriminar isolados microbianos e, ao mesmo tempo, estabelecer uma relação clonal entre linhagens relacionadas. Constituem, assim, métodos úteis para estudos epidemiológicos que visam prevenir e bloquear cadeias de transmissão de diversas doenças infecciosas (10,22). A aplicação desses métodos tem contribuído significativamente para uma maior compreensão dos casos e da transmissão da tuberculose em diversas regiões do mundo.…”