Saeed Zaker BOSTANABAD, Leonid P. TITOV, Mohammad TAGHIKHANI, Ahmadreza BAHRMAND

Identification of mutations in the rpoB encoding the RNA polymerase beta subunit in rifampicine-resistant Mycobacterium tuberculosis strains from Iran

Bu çalışmanın amacı; İranlı hastalardan izole edilen Mycobacterium tuberculosis suşlarında rpoB mutasyonlarının sıklığı, lokalizasyonu ve tiplerini belirlemekti. Tüberküloz şüphesi olan 91 hastadan balgam örnekleri alındı. M. tuberculosis olarak tanımlananların 34 (%87)’ünde Rif-r izole edildi. Polimeraz zincir reaksiyonu (PCR) amplifikasyonu ve DNA sekanslama metotları kullanıldı. rpoB geni 411 bp fragmanları sekanslandı ve 81 bp bölgelerinin mutasyonları incelendi. Yirmi dokuz RIF-r MBT’de (%85) 60 mutasyon ve 13 mikrodelesyon saptandı. Altmış mutasyon içinde altı sessiz ve 54 missens mutasyon belirlendi. Missens mutasyonlar 23 tip aminoasit değişikliğini ortaya çıkardı. Beş RIF-r MBT izolatında (%15) rpoB geni core bölgesinde hiçbir mutasyon saptanmadı. Tüm sessiz mutasyonların 507 numaralı kodonda yerleştiği belirlendi. İranlı suşlarda saptanan mutasyonlar en sık 523 ve 526 numaralı kodonlardaydı. Beş yüz yirmi altı numaralı kodonda 5 allel ve 507, 508 ve 513 numaralı kodonların her birinde üç allelde tripletler bulundu. Altı (%19) suşta 526, 510 numaralı kodonlarda tekli mutasyonlar varken, geri kalan 23 (%69) izolatta multipl mutasyonlar: ikili 11 (%34), üçlü 7 (%22), dörtlü 1 (%3) ve beşli 4 (%12) olduğu belirlendi.

İran’da rifampisine dirençli Mycobacterium tuberculosis suşlarında rpoB kodlayan RNA polymerase beta subunit mutasyonlarının tanımlanması

The aim of this study was to investigate the frequency, location and type of rpoB mutations in Mycobacterium tuberculosis isolated from patients in Iran. 91 sputum were collected from suspected tuberculosis patients, 34 Rif-r isolates (87%) were identified as M. tuberculosis. Polymerase chain reaction (PCR) amplification and DNA sequencing methods were performed. 411 bp fragments of rpoB gene were sequenced and mutations in 81 bp regions were analyzed. 60 mutations and 13 micro deletions were identified in 29 RIF-r MBT (85%). Among 60 mutations, 6 silent and 54 missense were identified. Missense mutations produced 23 types of amino acid substitutions. In 5 RIF-r MBT isolates (15%) no mutations were found in the core region of the rpoB gene. All silent mutations were localized in codon 507. Most frequent mutations detected from Iranian strains were in codons 523 and 526. Five alleles in codon 526 and 3 alleles in triplets in each codons 507, 508, 513 were found. 6 (19%) strains harboured single mutations 6 (18%) placed in codons 526, 510 while the rest of isolates 23 (69%) had multiple mutations: Double 11 (34%), triple 7 (22%), and quartile mutations 1 (3%) and 4 (12%) of strains harboured 5 mutations respectively.

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