AbstractDetermination of Coherency Between Polymerase Chain Reaction-Restriction Fragment Lenght Polymorphism Technique and Conventional Methods for the Identification of Mycobacteria at the Species Level Objective: There are still various problems at diagnosis and identification of tuberculosis. The aim of this study was to determine the coherency between molecular biologic and classical biochemical methods for the identification of Mycobacterium at the species level. Method: While nitrate, catalase and niacin tests were used for biochemical identification, Polymerase Chain Reaction-Restriction Fragment Length Polymorphism technique was used as molecular biologic method for isolated Mycobacterium spp. Results: It was detected that 30% of samples were positive with acid-fast bacilli smear before cultivation. While 43% of samples were culture positive by cultivation 30% of samples were positive for acid-fast bacilli smear and culture at the same time. 7 isolates (16.3%) were identified as Mycobacteria other than tuberculosis and 36 (83.7%) isolates were identified as Mycobacterium tuberculosis complex with Restriction Fragment Length Polymorphism technique. According to the biochemical identification, 40% of isolates had low activity for catalase test, 41% of isolates were positive for nitrate test, 30% of isolates were positive for niacin test. Conclusion: Coherency between Polymerase Chain Reaction-Restriction Fragment Length Polymorphism and conventional methods was evaluated by kappa statistics in our study. Coherency between two methods was determinated as middle degree. Disadvantages of Polymerase Chain Reaction-Restriction Fragment Length Polymorphism technique are false positive and negative results which can occur occasionally. We determined that classical and molecular techniques must be evaluated with clinical features at diagnosis of tuberculosis.
Mycobacterium tuberculosis complex Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) identification.
Amaç: Günümüzde tüberkülozun laboratuvar tanısında ve identifikasyonunda karşılaşılan çeşitli zorluklar mevcuttur. Bu çalışmada mikobakteri identifikasyonunda klasik biyokimyasal yöntemler ile moleküler yöntemler arasındaki uyumun belirlenmesi amaçlanmıştır. Yöntem: Biyokimyasal identifikasyonda, nitrat, katalaz ve niasin testleri, moleküler biyolojik teknik olarak Polimeraz Zincir Reaksiyonu-Parça Uzunluk Polimorfizmi yöntemi kullanılarak, Mycobacterium spp. olarak izole edilen mikobakterilerin identifikasyonu yapılmıştır. Bulgular: Örneklerin ekim öncesi yapılan Ehrlich Ziehl Neelsen direkt incelemelerinde aside dirençli basil örneklerin 30'unda (%30) saptanmıştır. Örneklerin 43\'ü (%43) kültür yöntemi ile pozitif bulunurken, Ehrlich Ziehl Neelsen ve kültürün aynı anda 30'u (%30) pozitif olarak belirlenmiştir. Polimeraz Zincir Reaksiyonu-Parça Uzunluk Polimorfizmi yöntemi ile Mycobacterium spp. olarak izole edilen 7 izolatın (%16.3) Nontüberküloz Mikobakteriler olduğu geri kalan 36 (%83.7) örneğin Mycobacterium tuberculosis complex olduğu tespit edildi. Klasik identifikasyon sonucunda 40 suşun katalaz testi düşük, 41 suşun nitratı pozitif bulunurken, 30 suşun niasin testi pozitif bulundu. Sonuç: Bu çalışmada biyokimyasal identifikasyonun moleküler yöntemle uyumu kapa istatistiği ile değerlendirildi. Bu iki yöntem arasındaki uyum orta derecede çıkmıştır. Polimeraz Zincir Reaksiyonu-Parça Uzunluk Polimorfizmi yönteminin en önemli dezavantajı zaman zaman yanlış pozitif ve negatif sonuçların ortaya çıkabilmesidir. Bu nedenle tüberkülozun laboratuvar tanısında klasik ve moleküler yöntemlerin sonuçları değerlendirilirken mutlaka klinik ön tanı ile birlikte değerlendirilmesi gerektiği düşüncesindeyiz.
Mycobacterium tuberculosis complex Polimeraz Zincir Reaksiyonu-Parça Uzunluk Polimorfizmi tekniği identifikasyon
Primary Language | Turkish |
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Journal Section | Articles |
Authors | |
Publication Date | September 1, 2008 |
Submission Date | June 13, 2014 |
Published in Issue | Year 2008 Volume: 1 Issue: 3 |
MEU Journal of Health Sciences Assoc was began to the publishing process in 2008 under the supervision of Assoc. Prof. Gönül Aslan, Editor-in-Chief, and affiliated to Mersin University Institute of Health Sciences. In March 2015, Prof. Dr. Caferi Tayyar Şaşmaz undertook the Editor-in Chief position and since then he has been in charge.
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