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Validation of Real Time PCR (TaqMan) Approach in Diagnosing Patients with Tuberculosis in Northeast of Iran
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| Author:
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MALIHEH BEHBODI GRO, MEHRI HOSSEINI, LEILA MANZOURI, ASGHAR SHARIFI, SEYED ABDOLRAHIM REZAEE, OWRANG EILAMI, SEYED REZA KHOSRAVANI, SEYED ABDOLMAJID KHOSRAVANI
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| Abstract:
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Background and objective: Accurate methods are needed for the better detection of tuberculosis (TB).
This study compared different laboratory tests and evaluated IS6110- Taqman system PCR for the detection of
mycobacterium tuberculosis (MTB) among suspected TB patients.
Method and material: Pulmonary and extra pulmonary samples were obtained from all patients with
symptoms similar to TB referred to hospitals affiliated of Mashhad University of Medical Sciences. Their
demographic information (age, type of sample, sex) and type of preparation were recorded. Samples were then
evaluated for smear and culture for positive or negative. Subsequently, DNA was extracted using DNA
isolation kit (Qiagen) for molecular analysis, then primers and probes were designed using Beacon designer
software and optimized by Real-time PCR. In this study, Real-time PCR Taqman probe for IS6110 fragment
was used to diagnose tuberculosis. All sputum samples were analyzed by smear microscopy, culture and realtime PCR with a view to detecting M. tuberculosis. Then data were analyzed using SPSS 21 software, through
descriptive and inferential statistics at a 95% confidence level. Also, for drawing the graphs we used Microsoft
Excel 2010. In this study, ethical considerations were respected.
Results: In this study 294 specimens were included, 20 positive smear samples and 213 negative. Of the 233
samples, 49 had positive TaqMan and 184 were negative. 61 samples were uncultivable, 1 of which was TaqMan
positive. Real-time PCR Taqman probe test was found to have sensitivity and specificity of 100% and 86.38%
respectively.
Conclusion: Since classical methods for detection of mycobacterium tuberculosis are time-consuming, so this
study was described as the fastest real-time PCR method. According to the results of this study, IS6110
Taqman probe Real-time PCR can be used as a standard method for diagnosis and confirmation of
tuberculosis.
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| Keyword:
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Mycobacterium tuberculosis, real-time PCR Taqman probe
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| EOI:
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| DOI:
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https://doi.org/10.31838/ijpr/2021.13.01.442
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| Download:
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