Mycobacterium tuberculosis kompleksinde izoniazid ve rifampisin direncinin real-time PCR ile hızlı tanısı
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2008
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info:eu-repo/semantics/openAccess
Özet
Otuz izoniazide dirençli, 30 rifampisine dirençli, 21 hem izoniazid hem de rifampisine dirençli ve kontrol grubu olarak her iki ilaca duyarlı 30 Mycobacterium tuberculosis suşu radyometrik BACTEC metodu kullanılarak belirlenmiştir. KatG and rpoB sensor probları kullanılarak, M.tuberculosis mutasyonlarının varlığı real-time PCR metodu ile incelenmiştir. İzoniazid için 17 izolatta prob Tm aralığı 66.98-67.70°C ve 6 izolatta prob Tm aralığı 68.19-68.90°C olarak belirlenmiştir. Toplam 23 izolatta dirence sebep olan mutasyonlar saptanmıştır. Rifampisin için, 9 izolatta prob Tm aralığı 65.3-66.70°C, 22 izolatta prob Tm aralığı 57.70-58.40°C ve 4 suşta bu Tm aralıklarının her ikisi gösterilmiştir. Toplam 27 suşta dirence sebep olan mutasyonlar belirlenmiştir. Real-time PCR metodu izoniazid direncini % 76.7 olarak belirlemiştir. BACTEC ve real-time PCR metodları arasında istatistiksel olarak anlamlı bir fark saptanmıştır (p=0.005). Real-time PCR ile rifampisin direnci % 90.0 olarak belirlenmiş, BACTEC ve real-time PCR arasında istatistiksel olarak anlamlı bir fark saptanmamıştır (p=0.076). Bu sonuçlar real-time PCR metodunun rutin laboratuvarda M.tuberculosis komplekste ilaç direncini belirlemede hızlı, basit ve güvenilir bir metod olduğunu göstermiştir.
Susceptibilities of 30 isoniazid-resistant, 30 rifampicin-resistant and 21 isoniazid and rifampisin-resistant Mycobacterium tuberculosis isolates as well as 30 isolates susceptible to the both drugs used as control group were determined by radyometric BACTEC method. Using katG and rpoB sensor probes, presence of mutations was investigated in M.tuberculosis by real-time PCR method. For isoniazid, prob Tm range of 66.98-67.70°C in 17 isolates and prob Tm range of 68.19-68.90°C in 6 isolates were found. Resistance-causing mutations were detected in a total of 23 isolates. For rifampicin, prob Tm range of 65.3-66.70°C in 9 isolates, prob Tm range of 57.70-58.40°C in 22 isolates and both of these Tm ranges in 4 isolates were determined. Mutations causing resistance were detected in a total of 27 isolates. By real-time PCR method isoniazid mutations were detected as 76.7 %. There was a statistically significant difference between BACTEC and real-time PCR methods (p=0.005). On the other hand, 90.0 % of rifampicin resistance were detected by real-time PCR, which indicated a statistically insignificant difference between BACTEC and real-time PCR (p=0.076). All these results showed that real-time PCR method is a fast, simple and accurate method for determining the drug resistance in M.tuberculosis complex in routine laboratories.
Susceptibilities of 30 isoniazid-resistant, 30 rifampicin-resistant and 21 isoniazid and rifampisin-resistant Mycobacterium tuberculosis isolates as well as 30 isolates susceptible to the both drugs used as control group were determined by radyometric BACTEC method. Using katG and rpoB sensor probes, presence of mutations was investigated in M.tuberculosis by real-time PCR method. For isoniazid, prob Tm range of 66.98-67.70°C in 17 isolates and prob Tm range of 68.19-68.90°C in 6 isolates were found. Resistance-causing mutations were detected in a total of 23 isolates. For rifampicin, prob Tm range of 65.3-66.70°C in 9 isolates, prob Tm range of 57.70-58.40°C in 22 isolates and both of these Tm ranges in 4 isolates were determined. Mutations causing resistance were detected in a total of 27 isolates. By real-time PCR method isoniazid mutations were detected as 76.7 %. There was a statistically significant difference between BACTEC and real-time PCR methods (p=0.005). On the other hand, 90.0 % of rifampicin resistance were detected by real-time PCR, which indicated a statistically insignificant difference between BACTEC and real-time PCR (p=0.076). All these results showed that real-time PCR method is a fast, simple and accurate method for determining the drug resistance in M.tuberculosis complex in routine laboratories.
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Farmakoloji ve Eczacılık
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22
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1
