Leonid P. TITOV, Manijeh KAHBAZI, Mohammad ARJOMANDZADEGAN, Larisa K. SURKOVA, Parisa FARNIA, Fatemeh SHEIKHOLESLAMI, Arezoo ESHGHINEJAD, Ali Asghar FARAZI, M. ESHRATI, Mana SHOJAPUR, Parviz OWLIA, Azam AHMADI

Determination of principal genotypic groups among susceptible, MDR and XDR clinical isolates of Mycobacterium tuberculosis in Belarus and Iran

Giriş: Mycobacterium tuberculosis complexin tüm üyeleri KatG463/GyrA95 polimorfizmi temelinde üç temel genetik gruptanbirinde yer alır.Materyal ve Metod: Beyaz Rusya ve İran (Tahran ve Markazi)ın değişik bölgelerinden kültürle doğrulanmış tüberkülozluhastalardan 50si duyarlı, 121i MDR (çoklu ilaç direnci) ve 31i XDR (yaygın ilaç direnci) toplam 202 M. tuberculosis izo-latı izole edildi. İzolatlar, sequencing ve PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism)ile incelendi ve KatG463 GyrA95 kodonlarda polimorfizm ile Sreevatsans patterni temelinde üç ana genetik gruba (PGG)ayrıldı.Bulgular: Beyaz Rusyadan MDR olarak tanımlanan 104 izolattan 57 (%54.8 ± 4.8)si, 30 (%28.8 ± 4.43)u ve 17 (%16.3 ±3.6)si sırasıyla PGG1, 2 ve 3te yer alıyordu (p< 0.05). Beyaz Rusyadan 31 XDR, 15 (%48.4)i, 12 (%38.7)si, 4 (%12.9)üsırasıyla PGG 1, 2 ve 3te olmak üzere benzer bir patterne sahipti. İran örneklerinden, Markazi izolatları (ilaca duyarlı) 12(%36.5), 15 (%45.5), 3 (%6), ve Tahran örnekleri (seçilmiş MDR): 9 (%53), 6 (%35.2), 2 (%11.8) (PGG 1, 2 ve 3, sırasıyla) pa-ternine sahipti. Cezaevinde yatan tüberkülozlu hastalarda yapılan bir çalışmada, izoniazide direnç ile PGG arasında ilişkibulunmadı, ancak saptanan izolatların çoğu PGG 1de yer alıyordu (%45.5 ± 10.9) (p< 0.05). Genel olarak, grup 1 izolatla-rı, MDR ve XDRde duyarlı türlere göre daha sıktı ve coğrafik bölge ile ilişki yoktu. Sonuç: Duyarlı türlerde dominant form grup 2 ve 3te bulunuyordu. PGG grupları, coğrafik orijinden çok izolatların dirençdurumu ile ilişkili gibi görünmektedir.

Determination of principal genotypic groups among susceptible, MDR and XDR clinical isolates of Mycobacterium tuberculosis in Belarus and Iran

Introduction: All members of the Mycobacterium tuberculosis complex were assigned to one of the three principle geneticgroups based on KatG463/GyrA95 polymorphism.Materials and Methods: A total of 202 isolates of M. tuberculosis consisting of 50 susceptible, 121 MDR (multidrug resis-tant) and 31 XDR (extensively drug resistant) isolated from culture-confirmed tuberculosis patients in different regions ofBelarus and Iran (Tehran and Markazi province). Isolates were screened by sequencing and polymerase chain reaction rest-riction fragment length polymorphism (RFLP) assay, and were further divided into three principal genetic groups (PGG),based on Sreevatsan s pattern as polymorphisms in KatG463/GyrA95 codons.Results: Among the 104 isolates, characterized as MDR from Belarus, 57 (54.8 ± 4.8%), 30 (28.8 ± 4.43%), 17 (16.3 ± 3.6),belonged to PGG 1, 2, and 3, respectively (p< 0.05). Thirty one XDR isolates from Belarus had a similar pattern as 15(48.4%), 12 (38.7%), 4 (12.9%) PGG 1, 2, and 3, respectively. From Iranian samples, Markazi isolates (susceptible todrugs) had a pattern as 12 (36.5%), 15 (45.5%), 3 (6%), and Tehran samples were (selected MDR): 9 (53%), 6 (35.2%), 2(11.8%) (PGG 1, 2, and 3, respectively). In a study of tuberculosis patients, who were in prison, no relation was foundbetween PGG and resistance to isoniazid, but most of the identified isolates belonged to PGG 1 (45.5 ± 10.9%) (p< 0.05).Overall, the group 1 isolates showed more frequency in MDR and XDR rather than susceptible strains, and there aren tany relations to geographic region.Conclusion: In susceptible strains, dominant forms were belonged to groups 2 and 3. It seems that PGG typing is closelyrelated to resistance status of isolates rather than geographic origin.

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1.Palomino JC, Leao SC, Ritacco V (eds). Tuberculosis from ba- sic science to patient care (electronic only). 2007; 620-87.
2. World Health Organization. Laboratory services in tuberculo- sis control. Part III. Culture 1998: 77.
3. World Health Organization, Global Health Atlas, Country Pro- files on Tuberculosis. Geneva, Switzerland. 2006.
4. World Health Organization Stop TB Department. Frequently asked questions: XDR-TB. 2006.
5. Setareh M, Titov LP, Surkova LK. High level association of mu- tation in KatG315 with MDR and XDR clinical isolates of Mycobacterium tuberculosis in Belarus; Acta Microbiologica et Immunologica Hungarica, 2009; 56: 313-25.
6. Arjomandzadegan M, Owlia P, Ranjbar R, Farazi AA, Sofian M, Sadrnia M, et al. Prevalence of mutations at codon 463 of katg gene in MDR and XDR clinical isolates of mycobacterium tuberculosis in Belarus and application of the method in rapid diagnosis; Acta Microbiologica et Immunologica Hungarica, 2011; 58: 51-63.
7. Sreevatsan S, Pan X, Stockbauer KE, Connell ND, Kreiswirth BN, Whittam TS, et al. Restricted structural gene polymorp- hism in the Mycobacterium tuberculosis complex indicates evolutionarily recent global dissemination. Proc Natl Acad Sci USA 1997; 94: 9869-74.
8. Cockerill III FR, Uhl JR, Temesgen Z, Zhang Y, Stockman L, Ro- berts GD, et al. Rapid identification of a point mutation of the Mycobacterium tuberculosis catalase-peroxidase (katG) gene associated with isoniazid resistance. J Infect Dis 1995; 171: 240-5.
9. Dolzani L, Rosato M, Sartori B, Banfi E, Lagatolla C, Predomi- nato M, et al. Mycobacterium tuberculosis isolates belonging to katG gyrA group 2 are associated with clustered cases of tu- berculosis in Italian patients. J Med Microbiol 2004; 53: 155-9.
10. Rhee JT, Piatek AS, Small PM, Harris LM, Chaparro SV, Kra- mer FR, et al. Molecular epidemiologic evaluation of transmis- sibility and virulence of Mycobacterium tuberculosis. J Clin Microbiol 1999; 37: 1764-70.
11. Rindi L, Lari N, Bonanni D, Garzelli C. Detection of Mycobac- terium tuberculosis genotypic groups by a duplex real-time PCR targeting the katG and gyrA genes. J Microbiol Meth 2004; 59: 283-7.
12. Soini H, Pan X, Amin A, Graviss EA, Siddiqui A, Musser JM. Characterization of Mycobacterium tuberculosis isolates from patients in Houston, Texas, by spoligotyping. J Clin Microbiol 2000; 38: 669-76.
13. Bifani PJ, Mathema B, Kurepina NE, Kreiswirth BN. Global dissemination of the Mycobacterium tuberculosis W Beijing fa- mily strains. Trends Microbiol 2002; 10: 45-52.
14. van Doorn HR, Kuijper EJ, van der Ende A, Welten AG, van Soolingen D, de Haas PE, et al. The susceptibility of Mycobac- terium tuberculosis to isoniazid and the Arg-->Leu mutation at codon 463 of katG are not associated. J Clin Microbiol 2001; 39: 1591-4.
15. Kong Y, Cave MD, Zhang L, Foxman B, Marrs CF, Bates JH, et al. Association between Mycobacterium tuberculosis Be- ijing/W Lineage Strain Infection and Extrathoracic Tuberculo- sis: Insights from Epidemiologic and Clinical Characterization of the Three Principal Genetic Groups of M. tuberculosis Clini- cal Isolates. J Clinical Micro 2007; 45: 409-14.
16. Ioerger TR, Yicheng F. The non-clonality of drug resistance in Beijing genotype isolates of Mycobacterium tuberculosis from the Western Cape of South Africa. BMC Genomics, 2010; 11: 670.
17. Walsh PS, Metzger DA, Higuchi R. Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material. Biotechniques 1991; 10: 506-13.