Mycobacterium tuberculosis suşlarında anti-tüberkülo ilaçlara direncin moleküler yöntemlerle araştırılması
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Date
2016
Authors
Sağlam, Sezcan
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Journal ISSN
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Publisher
Uludağ Üniversitesi
Abstract
Tüberküloz modern tanı, tedavi ve kontrol yöntemlerine rağmen tüm dünyada önemli bir sağlık problemidir. Tanıda altın standart halen M. tuberculosis kültürüdür. M. tuberculosis kültürü, etkenin tanımlanması ve ilaç duyarlılığının çalışılması açısından önemlidir. Rifampin bakterilerde protein sentezini etkileyerek, etambutol ise arabinozil transferaz enzim inhibisyonu ile arabinogalaktan sentezini engelleyerek antimikobakteriyal etki gösteren ve tüberküloz tedavisinde kullanılan major antitüberküloz ilaçlardandır. Bu çalışmada fenotipik ilaç duyarlılık testi ile rifampin ve/veya etambutol direnci belirlenen izolatlarda, rpoB ve embB gen bölgelerindeki mutasyon varlığının ve mutasyon paternlerinin DNA dizi analizi ile araştırılması amaçlanmıştır. Çalışmamızda rifampin dirençli 7, etambutol dirençli 15, hem rifampin hem de etambutol dirençli 6 olmak üzere toplam 28 suş incelenmiştir. Rifampin dirençli 13 suşun 11'inde (%84,6) rpoB geninde mutasyon, etambutol dirençli 21 suşun ise 14'ünde (%66,6) embB geninde mutasyon saptanmıştır. RpoB mutasyonu saptanan 11 suşun 10'unda 531. kodonda yer alan Ser531Leu (TCG-TTG) mutasyonu bulunmaktadır. Bir suşta ise His526Asn (CAC-AAC) mutasyonu saptanmıştır. En sık saptanan embB mutasyonu 297. kodonda yer alan Ser297Ala (TCG-GCG) mutasyonudur ve 8 suşta saptanmıştır. Dört suşta ise 306. kodonda Met306Val (ATG-GTG) mutasyonu saptanmıştır. İkişer suşta Gly406Asp (GGC-GAC) ve Glu405Asp (GAG-GAC) mutasyonu 1 suşta ise Glu378Ala (GAG-GCG) saptanmıştır. Anahtar kelimeler: Mycobacterium tuberculosis, dizi analizi, rpoB, embB
Investigation of Drug Resistant Mycobacterium tuberculosis Strains with Molecular Methods Tuberculosis is an important public health problem in all over the world despite of modern diagnostic, treatment and control methods. The gold standard for diagnosis is still M. tuberculosis culture. M. tuberculosis culture is important for identification and drug susceptibility tests. Rifampin and ethambutol which have the antimycobacterial effects, are both of the major anti-tuberculosis drugs used in the treatment of tuberculosis, rifampin affects protein synthesis in bacteria and ethambutol also inhibits the synthesis arabinogalactan with arabinosyl transferase enzyme inhibition, respectively. This study aimed to investigate the presence of mutations in the embB and rpoB gene regions and mutation patterns by DNA sequence analysis on isolates which have determined rifampicin and/or ethambutol resistance with phenotypic drug susceptibility test. In this study, we examined the 28 stains which consist of 7 rifampin resistant strains, 15 ethambutol resistant strains, and 6 both rifampin and ethambutol resistant strains. It has been found that there have been mutations on rpoB gene regions of 11 of the 13 (84.6%) rifampin resistant strains and also on embB gene regions of 12 of 21 (66.6%) etambutol resistant strains. It was determined that the 10 of 11 strains identified in rpoB mutation have Ser531Leu (TCG-TTG) mutation which have located at codon 531 and the 1 of 11 strains has also His526Asn (CAC-AAC) mutation. Ser297Ala (TCG-GCG) mutation is the most frequent embB mutation which was found at codon 297 and it was determined on 8 strains. It was also detected that the 4 strains have Met306Val (ATG-GTG) mutation which was found at codon 306, 2 strains have Gly406Asp (GGC-GAC) and Glu405Asp (GAG-GAC) mutation and finally one strain have Glu378Ala (GAG-GCG) mutation. Key words: Mycobacterium tuberculosis, sequence analysis, rpoB, embB
Investigation of Drug Resistant Mycobacterium tuberculosis Strains with Molecular Methods Tuberculosis is an important public health problem in all over the world despite of modern diagnostic, treatment and control methods. The gold standard for diagnosis is still M. tuberculosis culture. M. tuberculosis culture is important for identification and drug susceptibility tests. Rifampin and ethambutol which have the antimycobacterial effects, are both of the major anti-tuberculosis drugs used in the treatment of tuberculosis, rifampin affects protein synthesis in bacteria and ethambutol also inhibits the synthesis arabinogalactan with arabinosyl transferase enzyme inhibition, respectively. This study aimed to investigate the presence of mutations in the embB and rpoB gene regions and mutation patterns by DNA sequence analysis on isolates which have determined rifampicin and/or ethambutol resistance with phenotypic drug susceptibility test. In this study, we examined the 28 stains which consist of 7 rifampin resistant strains, 15 ethambutol resistant strains, and 6 both rifampin and ethambutol resistant strains. It has been found that there have been mutations on rpoB gene regions of 11 of the 13 (84.6%) rifampin resistant strains and also on embB gene regions of 12 of 21 (66.6%) etambutol resistant strains. It was determined that the 10 of 11 strains identified in rpoB mutation have Ser531Leu (TCG-TTG) mutation which have located at codon 531 and the 1 of 11 strains has also His526Asn (CAC-AAC) mutation. Ser297Ala (TCG-GCG) mutation is the most frequent embB mutation which was found at codon 297 and it was determined on 8 strains. It was also detected that the 4 strains have Met306Val (ATG-GTG) mutation which was found at codon 306, 2 strains have Gly406Asp (GGC-GAC) and Glu405Asp (GAG-GAC) mutation and finally one strain have Glu378Ala (GAG-GCG) mutation. Key words: Mycobacterium tuberculosis, sequence analysis, rpoB, embB
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Keywords
Mycobacterium tuberculosis, Dizi analizi, rpoB, embB, Sequence analysis
Citation
Sağlam, S. (2016). Mycobacterium tuberculosis suşlarında anti-tüberkülo ilaçlara direncin moleküler yöntemlerle araştırılması. Yayınlanmamış uzmanlık tezi. Uludağ Üniversitesi Tıp Fakültesi.