Pulmoner ve ekstrapulmoner örneklerde tüberkülozun erken tanısı ve rifampisin direncinin tespiti için geneXpert MTB/RIF testinin değerlendirilmesi

Amaç: Mycobacterium tuberculosis kompleks’in çok ilaca dirençli suşlarının ortaya çıkması ve tüberkülozun yeniden önem kazanması, yeni tanı ve antibiyotik duyarlılık test yöntemlerinin geliştirilmesini teşvik etmiştir. Biz de çalışmamızda bu amaçla son zamanlarda geliştirilmiş bir tanı yöntemi olan GeneXpert MTB/RIF testini, klinik örneklerden M. tuberculosis kompleksin tanımlanması ve rifampisin direncinin saptanmasında değerlendirdik. Gereç ve Yöntem: Çalışmamızda klasik ve tam otomatize kültür yöntemleri ile GeneXpert MTB/RIF testi karşılaştırıldı. 389 adet çeşitli klinik örneklerin (171 adet pulmoner ve 218 adet ekstrapulmoner) kültürleri, BACTEC MGIT 960 kültür sistemi ve Löwenstein-Jensen besiyeri ile yapıldı. Aynı zamanda GeneXpert MTB/RIF sistemi ile bu klinik örnekler test edildi. Bulgular: GeneXpert MTB/RIF testi ile 23 örnek, Löwenstein-Jensen besiyeri ve BACTEC MGIT 960 kültür sistemi ile 22 hasta örneği pozitif olarak tespit edildi. İki M. tuberculosis kompleks suşu GeneXpert MTB/RIF testi ile başlangıçta rifampisin dirençli olarak belirlendi, ancak testin tekrarlanması ile iki suştan biri duyarlı olarak tespit edildi. Kültür sonuçlarına göre hem yayma negatif hem de pozitif pulmoner örnekler için GeneXpert MTB/RIF testinin duyarlılığı %100 (16/16) olarak bulundu. Testin ekstrapulmoner örnekler için duyarlılığı ve özgüllüğü, sırasıyla %83.3 ve %99.5 olarak belirlendi.Sonuç: GeneXpert MTB/RIF testinin, tüberkülozun erken tanısında ve çok ilaca dirençli suşların tespitindeki başarılı sonuçlarıyla tüberküloz laboratuvarlarında etkin bir şekilde kullanılabilecek otomatize moleküler bir tanı testi olduğu düşünülmektedir.

Evaluation of the geneXpert MTB/RIF assay for early diagnosis of tuberculosis and detection of rifampicin resistance in pulmonary and extrapulmonary specimens

Purpose: The emerging of multi-drug resistant strains of Mycobacterium tuberculosis complex and the resurgence of tuberculosis have forced new methods for diagnosis and antimicrobial susceptibility to be developed. In this study, we evaluated a recently marketed diagnostic technique, GeneXpert MTB/RIF test, in terms of identifying Mycobacterium tuberculosis complex and detecting rifampicin resistance from clinical samples. Material and Methods: In our study, GeneXpert MTB/RIF assay was compared with conventional and automated culture methods. 389 various clinical samples (171 pulmonary and 218 ekstrapulmonary samples) were cultured with BACTEC MGIT 960 culture system and Löwenstein-Jensen medium and were tested with GeneXpert MTB / RIF test. Results: 23 samples were positive for Mycobacterium tuberculosis complex by GeneXpert MTB/RIF test, but 22 samples were positive for Löwenstein-Jensen medium and BACTEC MGIT 960 culture system. 2 of 23 samples were detected rifampicin resistant at the first study, but re-studying of these samples resulted as rifampicin resistant for only one sample. According to culture findings of all pulmonary samples, the sensitivity and specificity for GeneXpert MTB / RIF test were both 100% (16/16). The sensitivity and specificity for extrapulmonary samples were 83.3% and 99.5%, respectively.Conclusion: GeneXpert MTB / RIF test is an effectual automated molecular diagnostic technique with its successful and reliable performance in early diagnosis of tuberculosis and detecting multi-drug resistant strains.

___

  • World Health Organization (WHO). Global Tuberculosis Report 2015. WHO/HTM/TB/2015.22, 2015.
  • Hillemann D, Rüsch-Gerdes S, Boehme C, Richter E. Rapid molecular detection of extrapulmonary tuberculosis by the automated GeneXpert MTB/RIF System. J Clin Microbiol. 2011;49:1202-5.
  • Zignol M, van Gemert W, Falzon D, Jaramillo E, Blanc L, Raviglione M. Modernizing surveillance of antituberculosis drug resistance: from special surveys to routine testing. Clin Infect Dis. 2011;52:901-6.
  • Zeka A, Taşbakan S, Çavuşoğu C. Evaluation of the GeneXpert MTB/RIF assay for rapid diagnosis of tuberculosis and detection of rifampin resistance in pulmonary and extrapulmonary specimens. J Clin Microbiol. 2011;49:4138–41.
  • Moure R, Muñoz L, Torres M, Santin M, Martín R, Alcaide F. Rapid detection of mycobacterium tuberculosis complex and rifampin resistance in smear-negative clinical samples by use of an integrated real-time PCR method. J Clin Microbiol. 2011;49:1137–9.
  • Bunsow E, Ruiz-Serrano MJ, LópezRoa P, Kestler M, Viedma DG, Bouza E. Evaluation of GeneXpert MTB/RIF for the detection of Mycobacterium tuberculosis and resistance to rifampin in clinical specimens. J Infect. 2013:68:338-43.
  • Alvarez-Uria G, Azcona JM, Midde M, Naik PK, Reddy S, Reddy R. Rapid diagnosis of pulmonary and extrapulmonary tuberculosis in HIV-infected patients. comparison of LED fluorescent microscopy and the GeneXpert MTB/RIF assay in a district hospital in India. Tuberc Res Treat. 2012;2012:932862.
  • Cepheid® 2015. Xpert® MTB/RIF Dx System Operator Manual. Cepheid, Sunnyvale, CA, ABD
  • Lawn, S. D.; Nicol, M.P. Xpert® MTB/RIF assay: development, evaluation and implementation of a new rapid molecular diagnostic for tuberculosis and rifampicin resistance. Future microbial. 2011,6:1067-82.
  • Kalokhe, A.S., Shafiq, M., Lee, J.C. et al., Multidrugresistant tuberculosis drug susceptibility and molecular diagnostic testing: a review of the literature. Am J Med Sci. 2013;345:143–8.
  • Boehme CC, Nabeta P, Hillemann D, Nicol MP, Shenai S, Krapp F et al. Rapid molecular detection of tuberculosis and rifampin resistance. N Engl J Med. 2010;363:1005-15.
  • Lawn SD, Nicol MP. GeneXpert® MTB/RIF assay: development, evaluation and implementation of a new rapid molecular diagnostic for tuberculosis and rifampicin resistance. Future Microbiol. 2011;6:1067–82.
  • Marlowe EM, Novak-Weekley SM, Cumpio J, Sharp SE, Momeny MA, Babst A et al. Evaluation of the CepheidGeneXpert MTB/RIF assay for direct detection of mycobacterium tuberculosis complex in respiratory specimens. J Clin Microbiol. 2011;49:1621–3.
  • Özkütük N, Sürücüoğlu S. Orta prevalanslı bölgede akciğer ve akciğer dışı tüberküloz tanısında GeneXpert MTB/RIF testinin değerlendirilmesi. Mikrobiyol Bul. 2014;48:223-32.
  • Çiftçi IH, Aslan MH, Aşık G. Evaluation of GeneXpert MTB/RIF results for the detection mycobacterium tuberculosis in clinical samples. Mikrobiyol Bul. 2011;45:43-7.
  • Walusimbi S, Bwanga F, De Costa A, Haile M, Joloba M, Hoffner S. Meta-analysis to compare the accuracy of GeneXpert, MODS and the WHO 2007 algorithm for diagnosis of smear-negative pulmonary tuberculosis. BMC Infect Dis. 2013;13:507.
  • Chang K, Lu W, Wang J, Zhang K, Jia S, Li F, Deng S, Chen M. Rapid and effective diagnos is of tuberculosis and rifampicin resistance with GeneXpert MTB/RIF assay: a meta-analysis. J Infect 2012;64:580-8.
  • Bodmer T, Ströhle A. Diagnosing pulmonary tuberculosis with the GeneXpert MTB/RIF Test. J Vis Exp. 2012;62:3547
  • Ioannidis P, Papaventsis D, Karabela S, Nikolaou S, Panagi M, Raftopoulou E et al. Cepheid GeneXpert MTB/RIF assay for mycobacterium tuberculosis detection and rifampin resistance identification in patients with substantial clinical indications of tuberculosis and smear-negative microscopy results. J Clin Microbiol. 2011;49:3068–70.
  • Zmak L, Jankovic M, Jankovic VK. Evaluation of GeneXpert MTB/RIF assay for rapid molecular diagnosis of tuberculosis in a two-year period in Croatia. Int J Mycobacteriol. 2013;2:179–82.
  • Baylan O. Çok ilaca dirençli tüberkülozdan sonra yaygın ilaca dirençli ve tüm ilaçlara dirençli tüberküloz formları: eski hastalığın yeni yüzleri. Mikrobiyol Bul. 2011;45:181-95.
  • Tahaoğlu K. çok ilaca dirençli tüberküloz (ÇİD-TB). In Tüberküloz (Eds Ş Özkara, Z Kılıçaslan):534-53. İstanbul, Aves Yayıncılık, 2010.
  • Türkiye Halk Sağlığı Kurumu. TBC Daire Başkanlığı. Verem Savaş Raporu. Ankara, THSK, 2013.