Characterisation of pks15/1 in clinical isolates of Mycobacterium tuberculosis from Mexico.

Tuberculosis (TB) is an infectocontagious respiratory disease caused by members of the Mycobacterium tuberculosis complex. A 7 base pair (bp) deletion in the locus polyketide synthase (pks)15/1 is described as polymorphic among members of the M. tuberculosis complex, enabling the identification of Euro-American, Indo-Oceanic and Asian lineages. The aim of this study was to characterise this locus in TB isolates from Mexico. One hundred twenty clinical isolates were recovered from the states of Veracruz and Estado de Mexico. We determined the nucleotide sequence of a ± 400 bp fragment of the locus pks15/1, while genotypic characterisation was performed by spoligotyping. One hundred and fifty isolates contained the 7 bp deletion, while five had the wild type locus. Lineages X (22%), LAM (18%) and T (17%) were the most frequent; only three (2%) of the isolates were identified as Beijing and two (1%) EAI-Manila. The wild type pks15/1 locus was observed in all Asian lineage isolates tested. Our results confirm the utility of locus pks15/1 as a molecular marker for identifying Asian lineages of the M. tuberculosis complex. This marker could be of great value in the epidemiological surveillance of TB, especially in countries like Mexico, where the prevalence of such lineages is unknown.

Characterisation of pks15/1 in clinical isolates of Mycobacterium tuberculosis from Mexico

Tuberculosis (TB) is an infectocontagious respiratory disease caused by members of the Mycobacterium tuberculosis complex. A 7 base pair (bp) deletion in the locus polyketide synthase (pks)15/1 is described as polymorphic among members of the M. tuberculosis complex, enabling the identification of Euro-American, Indo-Oceanic and Asian lineages. The aim of this study was to characterise this locus in TB isolates from Mexico. One hundred twenty clinical isolates were recovered from the states of Veracruz and Estado de Mexico. We determined the nucleotide sequence of a ± 400 bp fragment of the locus pks15/1, while genotypic characterisation was performed by spoligotyping. One hundred and fifty isolates contained the 7 bp deletion, while five had the wild type locus. Lineages X (22%), LAM (18%) and T (17%) were the most frequent; only three (2%) of the isolates were identified as Beijing and two (1%) EAI-Manila. The wild type pks15/1 locus was observed in all Asian lineage isolates tested. Our results confirm the utility of locus pks15/1 as a molecular marker for identifying Asian lineages of the M. tuberculosis complex. This marker could be of great value in the epidemiological surveillance of TB, especially in countries like Mexico, where the prevalence of such lineages is unknown.

rrs and rpsL mutations in streptomycin-resistant isolates of Mycobacterium tuberculosis from Mexico.

BACKGROUND/PURPOSE: Mutations in rpsL and rrs genes are associated with resistance to streptomycin in tuberculosis, but important geographical variation exists in these mutations. The goal of this study was to characterize these mutations in isolates of streptomycin-resistant mycobacteria originating from southeast Mexico. METHODS: Mycobacteria were isolated from patients with suspected drug-resistant tuberculosis. Susceptibility tests were carried out using the fluorometric method, and rrs and rpsL DNA sequencing was performed by capillary electrophoresis. RESULTS: Some 136 drug-resistant isolates were recovered, of which 91(67%) exhibited resistance to streptomycin. Mutations in rpsL were observed in 18 isolates (19%) in codons 43 (A→G, K/R, n = 12) and 88 (A→G, K/R, n = 4; A→C, K/Q, n = 2). Mutations in rrs were observed in 26 isolates (28%). These were at nucleotides 513 (A→C, n = 8) and 516 (C→T, n = 6), and six novel mutations at nucleotides 483 (A→T, n = 2), 485 (A→G, n = 2), 496 (G→A, n = 2), 795 (C→T, n = 6), 870 (C→T, n = 3), and 907 (A→C, n = 3), with some isolates showing more than one mutation. Finally, 47 (52%) of the isolates showed no mutation. CONCLUSION: The variety and presence or absence of the mutations found suggest the circulation of an important diversity of strains and the existence of additional mechanisms contributing to streptomycin resistance in the region.