Short Communication

Quantification of mycobacterial proteins in extrapulmonary tuberculosis cases by nano-based real-time immuno-PCR

Centre for Biotechnology, Maharshi Dayanand University, Rohtak, 124001, Haryana, India

Special Centre for Nano Science & Advanced Instrumentation Research Facility, Jawaharlal Nehru University, New Delhi, 110067, India

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Anam Rais

Special Centre for Nano Science & Advanced Instrumentation Research Facility, Jawaharlal Nehru University, New Delhi, 110067, India

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Abhishek Sheoran

Department of Statistics, Ramanujan College, University of Delhi, New Delhi, 110019, India

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Ekta Kamra

Centre for Biotechnology, Maharshi Dayanand University, Rohtak, 124001, Haryana, India

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Preeti Mor

Centre for Biotechnology, Maharshi Dayanand University, Rohtak, 124001, Haryana, India

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Aishwarya Soni

Centre for Biotechnology, Maharshi Dayanand University, Rohtak, 124001, Haryana, India

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Suman Sharma

Department of Pathology, University of Health Sciences, Rohtak, 124001, Haryana, India

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Promod K Mehta

*Author for correspondence: Tel.: +91 989 650 4193;

E-mail Address: pkmehta3@hotmail.com

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E-mail Address: promod_fahs@sgtuniversity.org

https://orcid.org/0000-0002-9274-4028

Centre for Biotechnology, Maharshi Dayanand University, Rohtak, 124001, Haryana, India

Present address: Microbiology Department, Faculty of Allied Health Sciences, Shree Guru Gobind Singh Tricentenary University, Gurugram, 122505, Haryana, India

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Published Online:4 Aug 2023https://doi.org/10.2217/fmb-2023-0054

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Abstract

Aim: Diagnosis of extrapulmonary tuberculosis (EPTB) is difficult, and a rapid and dependable diagnostic test is urgently needed. Methods: A nano-based assay, SYBR Green magnetic bead-coupled gold nanoparticle-based real-time immuno-polymerase chain reaction (MB-AuNP-RT-I-PCR) was studied for the quantitative detection of Mycobacterium tuberculosis MPT-64+CFP-10 proteins in clinically suspected EPTB patients. Results: A wide range (270 fg/ml–9.9 ng/ml) of MPT-64+CFP-10 was quantified by MB-AuNP-RT-I-PCR in EPTB cases, whereas magneto-ELISA demonstrated a narrow range (1.8–10 ng/ml). Furthermore, high sensitivity (88.2%) and specificity (100%) were attained by MB-AuNP-RT-I-PCR in EPTB (n = 51) and non-TB control (n = 49) subjects, respectively. Both MB-AuNP-I-PCR/magneto-ELISA exhibited significantly lower (p < 0.05–0.01) sensitivities than MB-AuNP-RT-I-PCR. Conclusion: The MB-AuNP-RT-I-PCR described herein shows good diagnostic accuracy, which may translate into a credible diagnostic kit.

Plain language summary

Extrapulmonary tuberculosis (EPTB) is a type of tuberculosis disease caused by the bacteria Mycobacterium tuberculosis (Mtb) that affect other regions of the body, rather than the lungs. Detecting EPTB is difficult, and a fast and reliable test is needed. This study developed a test based on a small particle, known as a nanoparticle, to identify Mtb in people with EPTB. The test shows good accuracy and could be used for routine testing.

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Papers of special note have been highlighted as: • of interest; •• of considerable interest

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Vol. 18, No. 12

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History

Received 7 March 2023

Accepted 18 May 2023

Published online 4 August 2023

Published in print August 2023

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Supplementary data

To view the supplementary data that accompany this paper please visit the journal website at: www.futuremedicine.com/doi/suppl/10.2217/fmb-2023-0054

Acknowledgments

The authors acknowledge Meenakshi Chauhan, Devendra Pawar and Sanjeev Parshad, University of Health Sciences, Rohtak, India, for providing the clinical specimens of EPTB patients.

Financial & competing interests disclosure

PK Mehta and B Dahiya acknowledge the Council of Scientific and Industrial Research, New Delhi, India, for awarding the Emeritus Scientist Fellowship and Research Associate Fellowship, respectively. A Rais gratefully acknowledges Indian Council of Medical Research, New Delhi, India, for the award of a senior research fellowship. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

Clinical samples were obtained after taking written informed consent from the patients visiting the hospital of University of Health Sciences, Rohtak, India (from June 2019 to December, 2021). This work was approved by the Institutional Human Ethics Committees (IHEC/19/02; IHEC/19/03; HEC/2021/306 and HEC/2021/307) of Maharshi Dayanand University, Rohtak, India.

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