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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Pasteur Institute of Iran
2023-06-01
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| 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. |
| format | Article |
| id | doaj-art-5da989327c0f429fa44bbc0818a09205 |
| institution | Kabale University |
| issn | 2345-5349 2345-5330 |
| language | English |
| publishDate | 2023-06-01 |
| publisher | Pasteur Institute of Iran |
| record_format | Article |
| 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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