Volume 24, Issue 2 (2021)
mjms 2021, 24(2): 65-72 |
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Radaei T, Bakhshi B, Nikkhah M, Hamzehloo G. Evaluation of ku gene as a differential biomarker for rapid molecular detection of Mycobacterium tuberculosis complex using a real-time PCR assay. mjms 2021; 24 (2) :65-72
URL: http://mjms.modares.ac.ir/article-30-57247-en.html
URL: http://mjms.modares.ac.ir/article-30-57247-en.html
1- Ph.D. student in Medical Bacteriology, Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
2- Professor in Medical BacteriologyFaculty of Medical SciencesTarbiat Modares UniversityTehran, Iran , b.bakhshi@modares.ac.ir
3- Department of Nanobiotechnology, Faculty of Biological Sciences, Tarbiat Modares University Tehran, Iran
4- Tehran Regional Reference Laboratory for Tuberculosis, Tehran, Iran
2- Professor in Medical BacteriologyFaculty of Medical SciencesTarbiat Modares UniversityTehran, Iran , b.bakhshi@modares.ac.ir
3- Department of Nanobiotechnology, Faculty of Biological Sciences, Tarbiat Modares University Tehran, Iran
4- Tehran Regional Reference Laboratory for Tuberculosis, Tehran, Iran
Abstract: (1182 Views)
Purpose: currently TB diagnosis is limited by some major limitations in low-income and less experienced hospitals. Recently, it has been proposed that the ku gene of mycobacterial strains has the potential to be a highly specific and sensitive candidate biomarker for molecular detection of Mycobacterium tuberculosis (Mtb). This study was aimed to evaluate the specificity and sensitivity of a real-time PCR assay for detection of ku gene in Mtb complex to determine its applicability for Mtb identification.
Materials and methods: The identification of Mtb was confirmed using GeneXpert assay. Specific primers for ku gene were designed and the cycle threshold (Ct) value from the real-time PCR was used as a proxy measure of the cut-off point. Receiver operating characteristic (ROC) curve analysis was conducted to determine the diagnostic performance of ku gene in detecting Mtb directly from clinical specimens.
Results: ku amplification was interpreted as positive and negative based on Ct values, in which a value <38 was considered positive and a value >40 was considered negative. Our findings revealed that the ku gene was found to be distributed in all Mtb-positive samples. Of note, none of the Mtb-negative exhibited a specific signal in a maximum of 40 cycles.
Conclusions: The ku gene amplification using real-time PCR indicated high sensitivity and specificity for the detection of Mtb complex in sputum samples.
Materials and methods: The identification of Mtb was confirmed using GeneXpert assay. Specific primers for ku gene were designed and the cycle threshold (Ct) value from the real-time PCR was used as a proxy measure of the cut-off point. Receiver operating characteristic (ROC) curve analysis was conducted to determine the diagnostic performance of ku gene in detecting Mtb directly from clinical specimens.
Results: ku amplification was interpreted as positive and negative based on Ct values, in which a value <38 was considered positive and a value >40 was considered negative. Our findings revealed that the ku gene was found to be distributed in all Mtb-positive samples. Of note, none of the Mtb-negative exhibited a specific signal in a maximum of 40 cycles.
Conclusions: The ku gene amplification using real-time PCR indicated high sensitivity and specificity for the detection of Mtb complex in sputum samples.
Keywords: Mycobacterium tuberculosis, ku gene, biomarker, nucleic acid amplification tests (NAATs), GeneXpert MTB/RIF
Article Type: Original Research |
Subject:
Molecular Medicine
Received: 2021/11/19 | Accepted: 2022/02/12
Received: 2021/11/19 | Accepted: 2022/02/12
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