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Mycobacterium tuberculosis Suşlarınd a Klasik Kültür Metodu Ve Pcr-Heteroduplex Analizi İle Saptanan Rifampisin İlaç Duyarlılığının Karşılaştırılması

Comparison of Rifampicin Drug Susceptibility Determined by Conventional Culture Method and Pcr-Heteroduplex Analysis in Mycobacterium Tuberculosis Strains

Journal Name:

  • İnönü Üniversitesi Tıp Fakültesi Dergisi

Publication Year:

  • 2003

Key Words:

Keywords (Original Language):

Author NameUniversity of AuthorFaculty of Author
Abstract (2. Language): 
Resistance of Mycobacterium tuberculosis (MTB) to antituberculous drugs has been threatening the public-health grossly. Recently, because of high-level of resistance to rifampicin in MTB, treatment is becoming impossible. In this study, 127 rifampicin resistant and 33 rifampicin susceptible strains were randomly chosen from tuberculosis culture-positive isolates, isolated in the laboratory of the Ataturk Chest Diseases and Chest Surgery Center in Ankara, Turkey, over a 2-year period (1997 to 1998). Clinical samples collected from patients with suspected tuberculosis were studied for acid-fast bacilli stain (AFBS) and cultured for detecting MTB. PCR was used to amplify genetic loci associated with rifampicin resistance. The amplified 305-bp fragment of the rpoB gene was applied to Heteroduplex assay for rapid detection of rifampicin resistance phenotype in MTB. The 127 rifampicin-resistant and 33 rifampicin-susceptible isolates, selected according to conventional culture method, were tested again by PCR-Heteroduplex analysis. According to PCR-Heteroduplex analysis, 105 isolates of 160 M. tuberculosis were rifampicin resistant, 55 strains were rifampicin susceptible. Of 127 rifampicin-resistant isolates detected by conventional culture method, 101 had concordance with Heteroduplex analysis, whereas 26 isolates did not. While 4 strains were detected as rifampicin susceptible by conventional culture technique, they were rifampicin resistant by Heteroduplex analysis. It is thought that there might be a mutation out of fragment amplified, in 26 strains which were not observed as rifampicin resistant by Heteroduplex analysis; or a mistake in the result of resistance defined by classic culture method. In conclusion, however, PCR-Heteroduplex analysis is a rapid and reliable method to detect rifampicin resistance, although it is a relatively expensive technique.
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Abstract (Original Language): 
Mycobacterium tuberculosis'i n (MTB) antitüberküloz ilaçlara karşı direnç geliştirmesi büyük ölçüde halk sağlığını tehdit etmektedir. Son zamanlarda, MTB'deki rifampisin direncinin yüksek seviyede olması nedeniyle, tedavisi zorlaşmaktadır. Bu çalışmada, yaklaşık 2 yıl boyunca (1997-1998) Ankara, Atatürk Göğüs Hastalıkları ve Göğüs Cerrahisi Merkezi laboratuvarından izole edilen, kültürü pozitif izolatlardan 127 rifampisin dirençli ve 33 rifampisin duyarlı suşlar rasgele seçilmiştir. Tüberkülozdan şüphelenilen hastalardan toplanan klinik örneklerden asido resistan basil (ARB)çalışılmıştır ve MTB saptamak için kültüre alınmıştır. Rifampisin direnciyle ilgili gen bölgesini çoğaltmak için PCR kullanılmıştır. rpoB geninin çoğaltılan 305 baz çiftlik parçası için MTB'deki rifampisin direnç fenotipini hızlı saptamak amacıyla Heterodupleks analizi uygulanmıştır. Klasik kültür yöntemine göre seçilmiş 127 rifampisin dirençli ve 33 rifampisin duyarlı izolatlar tekrar PCR-Heterodupleks analizi ile test edilmişlerdir. PCR-Heterodupleks analizine göre 160 M. tuberculosis suşlarından 105'i rifampisin dirençli ve 55 suş ise rifampisin duyarlı bulunmuştur. Klasik kültür metoduyla saptanan 127 rifampisin dirençli izolattan 101'i Heterodupleks analizi ile uyumluydu, fakat 26 izolat uyum göstermemiştir. Dört suş klasik kültür yöntemiyle rifampisine duyarlı olarak saptandığı halde, heterodupleks analizi ile rifampisine dirençli bulunmuşlardır. Heterodupleks analizi ile rifampisin dirençli olarak gözlenmeyen 26 suşta çoğaltılan parçanın dışında bir mutasyon, yada klasik kültür metodu ile saptanan direnç sonuçlarında bir hata olabileceği düşünülmüştür. Sonuç olarak, yine de PCR-Heterodupleks analizi nispeten pahalı bir teknik olmasına rağmen, rifampisin direncini saptamak için hızlı ve güvenli bir metottur.
FULL TEXT (PDF): 
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  • 3
115-120
  • Turkish

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