Direct detection of Mycobacterium tuberculosis complex strains from sputum samples by nested PCR method
Journal Name:
- Gülhane Tıp Dergisi
Keywords (Original Language):
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Abstract (2. Language):
The purpose of this study was to
detect the presence of Mycobacterium tuberculosis complex strains
in sputum samples by IS6110-based
nested polymerase chain reaction
(PCR). The sputum samples of 120
patients with pulmonary tuberculosis were collected. After decontamination procedure, a portion of the
sputum samples was inoculated on
Löwenstein-Jensen medium and
then the samples were incubated at
35-37 ºC, for 4-8 weeks. Tuberculosis bacilles were identified in 105
out of 120 sputum cultures collected
from the patients with tuberculosis.
Therefore, the direct sputum samples and the DNAs extracted from
the culture of those 105 patients
were used for amplification of
IS6110 gene region by nested PCR. In
the result of the nested PCR
method, 78 (74.29%) of the direct
sputum samples were found to be
positive for M. tuberculosis complex, while 84 (80%) of the culture
samples were positive. When the
results of the direct sputum samples
were compared to the culture samples, the sensitivity and specificity
of nested PCR from direct sputum
samples were 92.8% (78/84), and
100% (21/21), respectively. This
study has shown that nested PCR
method is a practical, sensitive and
specific method for the detection of
M. tuberculosis complex strains from
sputum samples.
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Abstract (Original Language):
Bu çalışmanın amacı, balgam örneklerinde Mycobacterium tuberculosis komplex türlerinin IS6110 kökenli "nested polymerase chain reaction" (PCR) ile direkt olarak belirlenmesidir. Bu amaçla, çalışmada, akciğer tüberkülozu tanısı konmuş 120 hastadan alınan balgam örnekleri kullanıldı. Dekontaminasyon işlemini takiben, balgam örneklerinin bir kısmı Löwenstein-Jensen besiyerine ekildi ve örnekler 35-37 º C'de 4-8 hafta inkübe edildi. Kültür sonucunda, 105 örnekte tüberküloz basillerinin ürediği belirlendi. Bu nedenle, nested PCR metodunda 105 hastaya ait kültür ve direkt balgam örneklerinden elde edilen DNA'lar kullanıldı. Nested PCR sonucunda, toplam 105 direkt balgam örneğinin 78'inde (%74.29), kültür örneklerinin ise 84'ünde (%80) M. tuberculosis komplex türleri belirlendi. Kültürde gerçekleştirilen nested PCR ile kıyaslandığında direkt balgamda gerçekleştirilen nested PCR'ın duyarlılığı %92.8 (78/84), özgüllüğü ise %100 (21/21) olarak tespit edildi. Çalışmanın sonuçlarına göre, nested PCR metodunun direkt balgam örneklerinden M. tuberculosis komplex türlerinin belirlenmesinde pratik, özgül ve duyarlı bir metod olduğu kaydedilmiştir.
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