Derya GÜRSEL, Hale BOZKURT AKTOPRAK, Cengiz ÇAVUŞOĞLU

Evaluation of the genotype MTBDRplus assay for the diagnosis of tuberculosis and rapid detection of rifampin and isoniazid resistance in clinical specimens

Amaç: Bu çalışmada Genotype MTBDRplus testinin klinik örneklerden tüberküloz tanısı ve hızlı rifampin (RIF) ve izoniyazid (INH) saptanmasıdaki performansının belirlenmesi amaçlanmıştır. Yöntem ve gereç: Aralık 2007 ve 2009 tarihleri arasında Ege Üniversitesi Tıp Fakültesi Tıbbi Mikrobiyoloji Anabilim Dalı Mikobakteriyoloji Laboratuvarı’na rutin inceleme için gönderilen 57 hastaya ait toplam 90 klinik örnek (69 balgam örneği, 11 bronkoskopik aspirat, 5 bronkoalveoler lavaj, 4 derin trakeal aspirat, 1 lenf aspiratı) çalışmaya alındı. Bulgular: Toplam 90 klinik örneğin 80’inde (% 88,9) MTDRplus testiyle geçerli sonuç elde edildi. Direkt bakısı pozitif 82 örneğin 74’ünde (% 90,2) MTDRplus ile geçerli sonuç alınırken, direkt bakısı negatif sekiz örneğin ikisinde geçersiz sonuç alındı. MTBDRplus testiyle ilaç duyarlılık testi arasındaki genel uyumluluk oranı RİF direnci için % 97,5 (78/80) ve INH direncini için % 98,8 (79/80) olarak sapatandı. Sonuç: MTBDRplus klinik örneklerde ve kökenlerde RIF ve INH’a dirençli M. tuberculosis’in tanısında kullanılabilecek yararlı bir araçtır. Buna karşın test sonuçları mutlaka kültür ve ilaç duyarlılık testleriyle doğrulanmalıdır.

Klinik örneklerden tüberküloz tanısı ve hızlı rifampin ve izoniyazid direnci saptanması için geno type MTBDRPlus testinin değerlendirilmesi

Aim: To determine the performance of the Genotype MTBDRplus assay for diagnosis of tuberculosis and rapid detection of rifampin (RIF) and isoniazid (INH) resistance in clinical specimens. Materials and methods: A total of 90 clinical specimens of 57 patients (69 sputum samples, 11 bronchoscopic aspirates, 5 bronchoalveolar lavage, 4 deep tracheal aspirate, 1 lymph aspirate) sent to the Ege University Medical Faculty, Department of Medical Microbiology, Mycobacteriology Laboratory between December 2007 and 2009 during the clinical routine were included in the study. Results: Overall 80 valid results were obtained for 90 clinical specimens (88.9%) with MTBDRplus. While 74 of 82 (90.2%) smear positive specimens gave interpretable results by MTDRplus, 2 of 8 smear negative specimens gave invalid results. Th e overall rates of concordance between the results of the MTBDRplus assay and those of the drug susceptibility testing for the assessment of RIF and INH resistance were 97.5% (78/80) and 98.8% (79/80), respectively. Conclusion: Although the MTBDRplus assay could be a useful tool for rapid identifi cation of RIF- and INH-resistant Mycobacterium tuberculosis in both clinical samples and strains, the test results must always be confi rmed by culture and drug susceptibility testing.

PDF

___

1. Tuberculosis Service. 2009. Tuberculosis report in Turkey. Ministry of Health, Ankara, Turkey.
2. Cavusoglu C, Hilmioglu S, Guneri S, Bilgic A. Characterization of rpoB mutations in rifampin-resistant clinical isolates of Mycobacterium tuberculosis from Turkey by DNA sequencing and line probe assay. J Clin Microbiol 2002; 40: 4435-8.
3. Mani C, Selvakumar N, Narayanan S, Narayanan PR. Mutations in the rpoB gene of multidrug-resistant Mycobacterium tuberculosis clinical isolates from India. J Clin Microbiol 2001;39: 2987-90.
4. Telenti A, Imboden P, Marchesi F, Lowrie D, Cole S, Colston MJ et al. Detection of rifampicin-resistance mutations in Mycobacterium tuberculosis. Lancet 1993; 341: 647-50.
5. Zhang Y, Telenti A. Genetics of drug resistance in Mycobacterium tuberculosis. In: Hatfull GF, Jacobs WR Jr, Eds. Molecular genetics of mycobacteria. Washington DC: ASM Press; 2000. 235-251.
6. Hillemann D, Weizenegger M, Kubica T, Richter E, Niemann S. Use of the Genotype MTBDR assay for rapid detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis complex isolates. J Clin Microbiol 2005; 43: 3699-703.
7. Kim SY, Park YJ, Kim WI, Lee SH, Ludgerus Chang C, Kang SJ et al. Molecular analysis of isoniazid resistance in Mycobacterium tuberculosis isolates recovered from South Korea. Diagn Microbiol Infect Dis 2003; 47: 497-502.
8. Mokrousov I, Narvskaya O, Otten T, Limeschenko E, Steklova L, Vyshnevskiy B. High prevalence of KatG Ser315Th r substitution among isoniazid-resistant Mycobacterium tuberculosis clinical isolates from Northwestern Russia, 1996 to 2001. Antimicrob Agents Chemother 2002; 46: 1417-24.
9. Zhang M, Yue J, Yang YP, Zhang HM, Lei JQ, Jin RL et al. Detection of mutations associated with isoniazid resistance in Mycobacterium tuberculosis isolates from China. J Clin Microbiol 2005; 43: 5477-482.
10. Nikolayevsky V, Brown T, Balabanova Y, Ruddy M, Fedorin I, Drobniewski F. Detection of mutations associated with isoniazid and rifampin resistance in Mycobacterium tuberculosis isolates from Samara Region, Russian Federation. J Clin Microbiol 2004; 42: 4498-4502.
11. Cirillo DM, Piana F, Frisicale L, Quaranta M, Riccabone A, Penati V et al. Direct rapid diagnosis of rifampicin-resistant Mycobacterium tuberculosis infection in clinical samples by line probe assay (INNO LiPA Rif.TB). New Microbiol 2004; 27:221-7.
12. Makinen J, Marttila HJ, Marjamaki M, Viljanen MK, Soini H. Comparison of two commercially available DNA line probe assays for detection of multidrug-resistant Mycobacterium tuberculosis. J Clin Microbiol 2006; 44: 350-2.
13. Mani C, Selvakumar N, Narayanan S, Narayanan PR. Mutations in the rpoB gene of multidrug-resistant Mycobacterium tuberculosis clinical isolates from India. J Clin Microbiol 2001;39: 2987-90.
14. Kent PT, Kubica GP. Public health mycobacteriology. A guide for a level III laboratory. Centers for Disease Control, Atlanta, GA ; 1985.
15. Nikolayevskyy V, Balabanova Y, Simak T, Malomanova N, Fedorin I, Drobniewski F. Performance of the Genotype MTBDRplus assay in the diagnosis of tuberculosis and drug resistance in Samara, Russian Federation. BMC Clin Pathol 2009; 9: 2.
16. Lacoma A, Garcia-Sierra N, Prat C, Ruiz-Manzano J, Haba L, Rosés S et al. GenoType MTBDRplus assay for molecular detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis strains and clinical samples. J Clin Microbiol 2008; 46: 3660-7.
17. Hillemann D, Rüsch-Gerdes S, Richter E. Evaluation of the GenoType MTBDRplus assay for rifampin and isoniazid susceptibility testing of Mycobacterium tuberculosis strains and clinical specimens. J Clin Microbiol 2007; 45: 2635-40.
18. Miotto P, Piana F, Cirillo DM, Migliori GB. Genotype MTBDRplus: a further step toward rapid identifi cation of drug-resistant Mycobacterium tuberculosis. J Clin Microbiol 2008; 46: 393-4.
19. Causse M, Ruiz P, Gutierrez JB, Zerolo J, Casal M. Evaluation of new GenoType MTBDRplus for detection of resistance in cultures and direct specimens of Mycobacterium tuberculosis. Int J Tuberc Lung Dis 2008; 12: 1456-60.