Konvansiyonel ve moleküler yöntemler ile idrar örneklerinden Mycobacterium tuberculosıs izolasyonu ve tanımlanması
Küçük Resim Yok
Dosyalar
Tarih
2007
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Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Öz: Genitoüriner sistem tüberkülozunda, klinik ve radyolojik bulgulardaki çeşitlilik, eksik ya da yetersiz öykü ve basilin izolasyonundaki zorluklar nedeniyle tanı ve tedavi gecikebilmektedir. Bu çalışmada, üriner tüberküloz şüphesi ile laboratuvarımıza gönderilen örneklerden Mycobacterium tuberculosis'\n Ehrlich-Ziehl-Neelsen (EZN), kültür ve polimeraz zincir reaksiyonu ve restriksiyon analiz (PCR-RFLP) yöntemleri ile izolasyonu ve tanımlanması amaçlanmıştır. Ocak 2004-Temmuz 2006 tarihleri arasında 437 hastaya ait 1004 idrar örneği dekontaminasyon sonrası Lövvenstein-Jensen (LJ) ve/veya BACTEC 12B (Becton Dickinson, USA) besiyerlerine inoküle edilmiş ve hazırlanan preparatlar EZN yöntemi ile boyanarak değerlendirilmiştir. M.tuberculosis kompleks (MTC) ve tüberküloz dışı mikobakterilerin (MOTT) ayırımı nitro-alfa-asetilamino-beta-hidroksipropiofen (NAP) testi ile yapılmış, MTC izolatlarının primer antitüberküloz ilaçlara karşı duyarlılıkları BACTEC 460 TB (Becton Dickinson, USA) kültür sistemi ile araştırılmıştır. Kültürde Mycobacterium spp. izolasyonu yapıldığında, tanımlama için PCR-RFLP yöntemi kullanılmıştır. Hastaların 22'sinde (%5) konvansiyonel yöntemlerden (EZN, LJ ve/veya BACTEC) herhangi birisi ile pozitif sonuç alınmıştır. EZN boyama yöntemi ile 15 olguda aside dirençli basil (ARB) pozitifliği belirlenmiş, 17 örnekte ise kültürde üreme saptanmıştır. Bu hastaların 10'u her iki yöntemle de pozitif bulunurken, yedi hastada kültür pozitif EZN negatif, beş hastada da kültür negatif EZN pozitif olarak saptanmıştır. Bu beş hastanın mesane tümörü nedeniyle BCG tedavisi almakta olduğu belirlenmiştir. Kültürden izole edilen 17 susun 12'si (%70.5) MTC, beşi (%29.4) ise M.fortuitum olarak tanımlanmıştır. MTC izolatlarından sekizi (%66.7) tüm primer antitüberküloz ilaçlara karşı duyarlı, biri izoniazid (INH) ve etambutole (ETB), biri INH ve rifampisine (RIF), ikisi ise sadece INH'e karşı dirençli olarak tespit edilmiştir. Sonuç olarak, izole edilen mikobakterilerin tiplendirilebilmesi ve antitüberküloz duyarlılığın belirlenebilmesi için, kültürün mutlaka yapılmasının bir kez daha vurgulanması gerektiği kanısına varılmıştır.
Abstract: Genitourinary tuberculosis presents a challenge in diagnosis and treatment due to variations in clinical and radiological signs, insufficient patient history and difficulty in the isolation of the bacilli. The aim of this study was to isolate and identify Mycobacterium tuberculosis from the urine samples obtained from patients with suspected urinary tuberculosis admitted to our hospital by using Ehrlich-Ziehl-Neelsen (EZN), culture and polymerase chain reaction-restriction analysis (PCR-RFLP) methods. A total of 1004 urine samples collected from 437 patients who were admitted to our hospital between January 2004- July 2006, were inoculated on Lowenstein-Jensen (LJ) and/or BACTEC 12B (Becton Dickinson, USA) after decontamination and, direct preparations stained with EZN method were evaluated microscopically. M.tuberculosis complex (MTC) and mycobacteria other than tuberculosis (MOTT) were differentiated by nitro-alpha-acetylamino-beta-hydroxypropiophenone (NAP) test and the susceptibility testing for the MTC strains to primary antituberculosis drugs were performed by BACTEC 460 TB (Becton Dickinson, USA) system. PCR-RFLP method was performed for the identification of Mycobacterium spp. Twenty-two (5%) patients have yielded positive results by at least one of the conventional methods (EZN, LJ and/or BACTEC). Fifteen samples were positive for acido-resistant bacilli (ARB) by EZN method, and 17 samples were positive for mycobacterial growth in the cultures. Ten of 22 patients were found positive by both of the methods, while seven were culture positive but ARB negative and five were culture negative but ARB positive. These five patients received BCG treatment because of the presence of bladder tumor. Twelve (70.5%) of 17 strains isolated from culture were identified as MTC, while five (29.4%) were identified as M.fortuitum. Of 12 MTC isolates, eight (66.7%) were found susceptible to all of the antituberculosis agents, while one was found resistant to isoniazide (INH) and ethambutole (ETB), one was resistant to INH and rifampicin (RIF), and two were resistant to only INH. It is concluded that, in order to identify mycobacteria and to perform antituberculous susceptibility tests, cultivation of mycobacteria is a prerequisite.
Abstract: Genitourinary tuberculosis presents a challenge in diagnosis and treatment due to variations in clinical and radiological signs, insufficient patient history and difficulty in the isolation of the bacilli. The aim of this study was to isolate and identify Mycobacterium tuberculosis from the urine samples obtained from patients with suspected urinary tuberculosis admitted to our hospital by using Ehrlich-Ziehl-Neelsen (EZN), culture and polymerase chain reaction-restriction analysis (PCR-RFLP) methods. A total of 1004 urine samples collected from 437 patients who were admitted to our hospital between January 2004- July 2006, were inoculated on Lowenstein-Jensen (LJ) and/or BACTEC 12B (Becton Dickinson, USA) after decontamination and, direct preparations stained with EZN method were evaluated microscopically. M.tuberculosis complex (MTC) and mycobacteria other than tuberculosis (MOTT) were differentiated by nitro-alpha-acetylamino-beta-hydroxypropiophenone (NAP) test and the susceptibility testing for the MTC strains to primary antituberculosis drugs were performed by BACTEC 460 TB (Becton Dickinson, USA) system. PCR-RFLP method was performed for the identification of Mycobacterium spp. Twenty-two (5%) patients have yielded positive results by at least one of the conventional methods (EZN, LJ and/or BACTEC). Fifteen samples were positive for acido-resistant bacilli (ARB) by EZN method, and 17 samples were positive for mycobacterial growth in the cultures. Ten of 22 patients were found positive by both of the methods, while seven were culture positive but ARB negative and five were culture negative but ARB positive. These five patients received BCG treatment because of the presence of bladder tumor. Twelve (70.5%) of 17 strains isolated from culture were identified as MTC, while five (29.4%) were identified as M.fortuitum. Of 12 MTC isolates, eight (66.7%) were found susceptible to all of the antituberculosis agents, while one was found resistant to isoniazide (INH) and ethambutole (ETB), one was resistant to INH and rifampicin (RIF), and two were resistant to only INH. It is concluded that, in order to identify mycobacteria and to perform antituberculous susceptibility tests, cultivation of mycobacteria is a prerequisite.
Açıklama
Anahtar Kelimeler
Kaynak
Mikrobiyoloji Bülteni
WoS Q Değeri
Scopus Q Değeri
Cilt
41
Sayı
2
Künye
ASLAN G,DORUK E,EMEKDAŞ G,SERİN M. S,DİREKEL Ş,ABİHA G. B,DURMAZ R (2007). Konvansiyonel ve moleküler yöntemler ile idrar örneklerinden Mycobacterium tuberculosıs izolasyonu ve tanımlanması. Mikrobiyoloji Bülteni, 41(2), 185 - 192.
