Mohammad ARJOMANDZADEGAN, Somayeh GRAVAND

Analysis of rpsL and rrs genes mutations related to streptomycin resistance in Mdr and Xdr clinical isolates of Mycobacterium tuberculosis

Giriş: Streptomisin, çok ilaca dirençli tüberküloz tedavisinde en etkili ilaçlardan birisidir. Bakterisidal aminoglikozid bir antibiyotiktir ve direnç giderek artmaktadır. Streptomisinin etki mekanizması aminoaçil tRNA'nın uzama fazında "A"ya bağlanmasını engeller ve sonuç olarak bakteriyel protein sentezini durdurur. Bu çalışmada Mycobacterium tuberculosis'in klinik örneklerinde streptomisin direncinin hızlı incelemesi çalışılmıştır. Materyal ve Metod: Bu çalışmada balgam ve kültür pozitif 105 M. tuberculosis'ten 45 tane streptomisine dirençli ve duyarlı örnek seçilmiş ve rrs ve rpsL genlerindeki mutasyonlar açısından çalışılmıştır. Polimeraz zincir reaksiyonu (PCR) için kullanılan spesifik primerler rpsL 1, rpsL 2, rrsR ve Frrs olarak adlandırılmıştır. PCR ürünleri sekanslanmıştır.Bulgular: rpsL geni için 504 bp bandı ve rrs geni için 1027 bp bandı gösteren PCR ürünleri, uygun primerlerin seçimi ve uygun amplifikasyon programını sağlamıştır. Streptomisine dirençli 26 örnekten 26'sında spsL geninde mutasyon ve birinde rrs geninde değişklik vardı. Bu çalışmada streptomisine duyarlı 19 örnekte mutasyon saptanmadı.Sonuç: Streptomisin direncinden spsL geninde 43 ve 88 kodonlarda ve nadiren rrs genindeki mutasyonlar sorumludur.

Mdr ve Xdr Mycobacterium tuberculosis örneklerinden streptomisin direncinden sorumlu rpsL ve rrs gen mutasyonlarının analizi

Introduction: Streptomycin is a bactericidal and aminoglycoside antibiotic. It is one of the most effective drugs for treatment of multidrug Tuberculosis disease. Incidence of resistance is increasingly reported. Its action mechanism is by inhibition of binding aminoacyl tRNA to position "A" in elongation phase, which finally it causes to stop bacterial protein synthesis. In this study, resistance rapid investigation to streptomycin was conducted in clinical strains of Mycobacterium tuberculosis. Material and Method: In this study, among 105 strains of phlegm-positive and culture-positive Mycobacterium tuberculosis, 45 strains of resistant and sensitive to streptomycin were selected for possible mutations examination in genes rrs and rpsL. Specific primers that used for PCR were named rpsL 1, rpsL 2 and rrsR, Frrs. PCR products were sequenced. Results: PCR Products represents 504 bp band for gene rpsL and 1027 bp for gene rrs that shows proper selection of primers and determining an amplification appropriate program. From 26 resistant strains to streptomycin 26 strain have mutation in rpsL gene and 1 strain have alteration in rrs gene. In this study 19 strains were sensitive to streptomycin that have no mutation in these gene. Conclusions: Streptomycin resistance is mainly related to mutation at codons 43 and 88 "rpsL" gene and to a lesser extent "rrs" that are the greatest cause of drug resistance to streptomycin.

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