Leonid P. TITOV, Saeed Z. BOSTANABAD, Seyed Ali NOJOUMI, Esmaeil JABBARZADEH, Mehdi SHEKARABEI, Hassan HOSEINAEI, Mohammad K. RAHIMI, Mostafa GHALAMI, Shahin POURAZAR DIZJI, Evgeni R. SAGALCHYK

High level isoniazid resistance correlates with multiple mutation in the katG encoding catalase proxidase of pulmonary tuberculosis isolates from the frontier localities of Iran

Bu çalışmanın amacı, katG genindeki multipl mutasyonların, nükleotid değişikliklerinin önemi ve İran’ın farklı coğrafik bölgelerinden rastgele toplanan primer ve sekonder aktif pulmoner tüberkülozlu 42 hastanın balgamında izoniazide yüksek düzeyde dirençli Mycobacterium tuberculosis izolatları ile ilişkisini araştırmaktı. İlaç duyarlılık testi, CDC standart konvansiyonel metod kullanılarak yapıldı. DNA ekstraksiyonu, katG gen amplifikasyonu ve DNA sekans analizi yapıldı. İzolatların 34 (%80)’ünde katG geninde multipl mutasyon (2-5 mutasyon) saptandı. İzoniazide yüksek düzeyde dirençli (MİK μg/mL ≥ 5- 10) sekonder tüberkülozlu hastalarda daha sık olmak üzere, 315 (AGC→ACC), 316 (GGC→AGC), 309 (GGT→GTT) kodonlarında artmış predominant mutasyon ve nükleotid değişiklikleri saptandı. Ayrıca, sekonder infeksiyonlu hastalarda daha sık mutasyonu gösteren, 315, 316 ve 309 kodonlarında mutasyon kombinasyonları ve predominant nükleotid değişiklikleri gözlendi. Bu çalışmada, multipl mutasyonlu izolatların %62 (n= 21)’sinde, 315 (AGC→ACC), 316 (GGC→GTT), 309 (GGT→GGT) kodonlarında mutasyon kombinasyonları ile predominant nükleotid değişiklikleri olduğu bulundu ve bunun da izoniazide yüksek dirençli (≥ 5-10 μg/mL) sekonder infeksiyonlu hastaların izolatlarında daha sık olduğu saptandı.

İran sınır bölgesindeki pulmoner tüberküloz izolatlarında yüksek düzeyde izoniazid direnci katalaz-peroksidazı kodlayan katG’deki multipl mutasyonla koreledir

The aim of this study was to investigate the significance of multiple-mutations in the katG gene, predominant nucleotide changes and its correlation with high level of resistance to isoniazid in Mycobacterium tuberculosis isolates that were randomly collected from sputa of 42 patients with primary and secondary active pulmonary tuberculosis from different geographic regions of Iran. Drug susceptibility testing was determined using the CDC standard conventional proportional method. DNA extraction, katG gene amplification, and DNA sequencing analysis were performed. Thirty four (80%) isolates were found to have multiple-mutations (composed of 2-5 mutations) in the katG gene. Increased number of predominant mutations and nucleotide changes were demonstrated in codons 315 (AGC→ACC), 316 (GGC→AGC), 309 (GGT→GTT) with a higher frequency among patients bearing secondary tuberculosis infection with elevated levels of resistance to isoniazid (MIC ≥ 5-10 μg/mL). Furthermore, it was demonstrated that the combination of mutations with their predominant nucleotide changes were also observed in codons 315, 316, and 309 indicating higher frequencies of mutations among patients with secondary infection respectively. In this study, 62% (n= 21) of multi-mutated isolates found to have combination of mutations with predominant nucleotide changes in codons 315 (AGC→ACC), 316 (GGC→GTT), 309 (GGT→GGT), and also demonstrated to be more frequent in isolates of patients with secondary infections, bearing higher level of resistance to isoniazid (≥ 5-10 μg/mL).

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