Multiplex PCR targeting IS6110 and mpb64 Genes for Detecting Extra- Pulmonary Tuberculosis: A Cross-Sectional Study in a Tertiary Care Center, Kashmir, India

Introduction: Extra-pulmonary tuberculosis (EPTB) is a significant cause of morbidity, and early diagnosis is critical for improving patient outcomes. Conventional diagnostic methods for EPTB often require improvement, highlighting the need for more rapid and sensitive diagnostic procedures. In t...

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Main Authors: Dekyong Angmo1, Gulnaz Bashir2*, Abiroo Jan3, Mushtaq A. Khan4, Syed Besina Yasin5
Format: Article
Language:English
Published: Pasteur Institute of Iran 2023-06-01
Series:Journal of Medical Microbiology and Infectious Diseases
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Online Access:https://jommid.pasteur.ac.ir/article-1-498-en.html
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author Dekyong Angmo1
Gulnaz Bashir2*
Abiroo Jan3
Mushtaq A. Khan4
Syed Besina Yasin5
author_facet Dekyong Angmo1
Gulnaz Bashir2*
Abiroo Jan3
Mushtaq A. Khan4
Syed Besina Yasin5
author_sort Dekyong Angmo1
collection DOAJ
description Introduction: Extra-pulmonary tuberculosis (EPTB) is a significant cause of morbidity, and early diagnosis is critical for improving patient outcomes. Conventional diagnostic methods for EPTB often require improvement, highlighting the need for more rapid and sensitive diagnostic procedures. In this cross-sectional study, we aimed to evaluate the diagnostic usefulness of multiplex PCR (mPCR) using IS6110 and mpb64 as gene targets for detecting Mycobacterium tuberculosis in samples from suspected cases of EPTB. We compared the performance of mPCR with conventional methods, including Ziehl Neelsen (ZN) microscopy, culture in LJ media, and BacT/Alert system. Our study aimed to provide insight into the utility of mPCR and its different targets for diagnosing EPTB in our setting. Methods: We conducted a crosssectional survey of 250 non-repeat clinical samples from extrapulmonary sites to detect M. tuberculosis. Both conventional diagnostic methods, including ZN microscopy, culture in LJ media, and BacT/Alert system, and molecular methods, including multiplex PCR (mPCR) using IS6110 and mpb64 as gene targets, were performed on the samples. Of the 250 samples, results for all the diagnostic methods were available for 116 samples, which were included in the final analysis. The study population comprised 83 patients with suspected EPTB and 33 controls. Results: Among the 83 samples in the EPTB group, conventional diagnostic methods, including ZN microscopy, LJ culture, and BacT/Alert system, showed low positivity rates of 6.02%, 8.43%, and 15.66%, respectively. In contrast, multiplex PCR (mPCR) using IS6110 and mpb64 as gene targets showed a significantly higher positivity rate of 79.51%. The IS6110 gene was amplified in 79.51% of the samples, while mpb64 was amplified in 49.39%. Conclusion: Our study demonstrates that multiplex PCR (mPCR) using IS6110 and mpb64 as gene targets is a more sensitive diagnostic method for extra-pulmonary tuberculosis (EPTB) than conventional methods. Both IS6110 and mpb64 showed high sensitivity of 100%, but mpb64 was more specific when compared with the gold standard. Our findings suggest that mPCR, particularly with the inclusion of mpb64 as the target gene, may be a valuable tool for the early and accurate diagnosis of EPTB.
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publisher Pasteur Institute of Iran
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series Journal of Medical Microbiology and Infectious Diseases
spelling doaj-art-5da989327c0f429fa44bbc0818a092052025-08-20T03:53:21ZengPasteur Institute of IranJournal of Medical Microbiology and Infectious Diseases2345-53492345-53302023-06-01112788510.61186/JoMMID.11.2.78Multiplex PCR targeting IS6110 and mpb64 Genes for Detecting Extra- Pulmonary Tuberculosis: A Cross-Sectional Study in a Tertiary Care Center, Kashmir, IndiaDekyong Angmo10https://orcid.org/0009-0006-2919-5413Gulnaz Bashir2*1https://orcid.org/0000-0002-8048-0854Abiroo Jan32https://orcid.org/0000-0002-3150-2906Mushtaq A. Khan43https://orcid.org/0000-0002-1257-6002Syed Besina Yasin54https://orcid.org/0000-0001-9886-11131Department of Microbiology, Government Medical College, Srinagar. Jammu and Kashmir, India2Department of Microbiology, Sher-i-Kashmir Institute of Medical Sciences, Soura, Srinagar, Jammu and Kashmir, India3Department of Microbiology, Government Medical College, Anantnag, Jammu and Kashmir, India4Department of Gastroenterology, Sher-i-Kashmir Institute of Medical Sciences, Soura, Srinagar, Jammu and Kashmir, India5Department of Pathology, Sher-i-Kashmir Institute of Medical Sciences, Soura, Srinagar, Jammu and Kashmir, IndiaIntroduction: Extra-pulmonary tuberculosis (EPTB) is a significant cause of morbidity, and early diagnosis is critical for improving patient outcomes. Conventional diagnostic methods for EPTB often require improvement, highlighting the need for more rapid and sensitive diagnostic procedures. In this cross-sectional study, we aimed to evaluate the diagnostic usefulness of multiplex PCR (mPCR) using IS6110 and mpb64 as gene targets for detecting Mycobacterium tuberculosis in samples from suspected cases of EPTB. We compared the performance of mPCR with conventional methods, including Ziehl Neelsen (ZN) microscopy, culture in LJ media, and BacT/Alert system. Our study aimed to provide insight into the utility of mPCR and its different targets for diagnosing EPTB in our setting. Methods: We conducted a crosssectional survey of 250 non-repeat clinical samples from extrapulmonary sites to detect M. tuberculosis. Both conventional diagnostic methods, including ZN microscopy, culture in LJ media, and BacT/Alert system, and molecular methods, including multiplex PCR (mPCR) using IS6110 and mpb64 as gene targets, were performed on the samples. Of the 250 samples, results for all the diagnostic methods were available for 116 samples, which were included in the final analysis. The study population comprised 83 patients with suspected EPTB and 33 controls. Results: Among the 83 samples in the EPTB group, conventional diagnostic methods, including ZN microscopy, LJ culture, and BacT/Alert system, showed low positivity rates of 6.02%, 8.43%, and 15.66%, respectively. In contrast, multiplex PCR (mPCR) using IS6110 and mpb64 as gene targets showed a significantly higher positivity rate of 79.51%. The IS6110 gene was amplified in 79.51% of the samples, while mpb64 was amplified in 49.39%. Conclusion: Our study demonstrates that multiplex PCR (mPCR) using IS6110 and mpb64 as gene targets is a more sensitive diagnostic method for extra-pulmonary tuberculosis (EPTB) than conventional methods. Both IS6110 and mpb64 showed high sensitivity of 100%, but mpb64 was more specific when compared with the gold standard. Our findings suggest that mPCR, particularly with the inclusion of mpb64 as the target gene, may be a valuable tool for the early and accurate diagnosis of EPTB.https://jommid.pasteur.ac.ir/article-1-498-en.htmleptbmpcrmpb64is6110tuberculosis (tb)diagnostic accuracysensitivityspecificity
spellingShingle Dekyong Angmo1
Gulnaz Bashir2*
Abiroo Jan3
Mushtaq A. Khan4
Syed Besina Yasin5
Multiplex PCR targeting IS6110 and mpb64 Genes for Detecting Extra- Pulmonary Tuberculosis: A Cross-Sectional Study in a Tertiary Care Center, Kashmir, India
Journal of Medical Microbiology and Infectious Diseases
eptb
mpcr
mpb64
is6110
tuberculosis (tb)
diagnostic accuracy
sensitivity
specificity
title Multiplex PCR targeting IS6110 and mpb64 Genes for Detecting Extra- Pulmonary Tuberculosis: A Cross-Sectional Study in a Tertiary Care Center, Kashmir, India
title_full Multiplex PCR targeting IS6110 and mpb64 Genes for Detecting Extra- Pulmonary Tuberculosis: A Cross-Sectional Study in a Tertiary Care Center, Kashmir, India
title_fullStr Multiplex PCR targeting IS6110 and mpb64 Genes for Detecting Extra- Pulmonary Tuberculosis: A Cross-Sectional Study in a Tertiary Care Center, Kashmir, India
title_full_unstemmed Multiplex PCR targeting IS6110 and mpb64 Genes for Detecting Extra- Pulmonary Tuberculosis: A Cross-Sectional Study in a Tertiary Care Center, Kashmir, India
title_short Multiplex PCR targeting IS6110 and mpb64 Genes for Detecting Extra- Pulmonary Tuberculosis: A Cross-Sectional Study in a Tertiary Care Center, Kashmir, India
title_sort multiplex pcr targeting is6110 and mpb64 genes for detecting extra pulmonary tuberculosis a cross sectional study in a tertiary care center kashmir india
topic eptb
mpcr
mpb64
is6110
tuberculosis (tb)
diagnostic accuracy
sensitivity
specificity
url https://jommid.pasteur.ac.ir/article-1-498-en.html
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