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Research Article

Quantitative detection of a cocktail of mycobacterial MPT64 and PstS1 in tuberculosis patients by real-time immuno-PCR

    Suman Sharma

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

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    ,
    Abhishek Sheoran

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

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    ,
    Krishna B Gupta

    Department of TB & Respiratory Medicine, University of Health Sciences, Rohtak-124001, India

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    ,
    Aparna Yadav

    Department of Microbiology, University of Health Sciences, Rohtak-124001, India

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    ,
    Mandira Varma-Basil

    Microbiology Department, Vallabhbhai Patel Chest Institute, University of Delhi, New Delhi-110007, India

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    ,
    Vishnubhatla Sreenivas

    Department of Biostatistics, All India Institute of Medical Sciences, New Delhi-110029, India

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    ,
    Dhruva Chaudhary

    Department of Pulmonary & Critical Care Medicine, University of Health Sciences, Rohtak-124001, India

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    &
    Promod K Mehta

    *Author for correspondence: Tel.: +91 989 650 4193; Fax: +91 126 227 4640;

    E-mail Address: pkmehta3@hotmail.com

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

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    Published Online:21 Jan 2019https://doi.org/10.2217/fmb-2018-0284

    Abstract

    Aim: There is an urgent need to design a reliable diagnostic test for tuberculosis (TB). Methods: Real-time immuno-PCR (RT-I-PCR) assay was devised for the quantitative detection of a cocktail of mycobacterial MPT64 (Rv1980c) and PstS1 (Rv0934) in TB patients. Results: A broad dynamic range of 0.95 pg/ml–95 ng/ml of MPT64+PstS1 was detected in TB patients. In smear-positive (n = 59) and smear-negative (n = 42) pulmonary TB cases, sensitivities of 93.2 and 83.3% were observed, respectively with 92.8% specificity, whereas a sensitivity of 77.9% and a specificity of 91.3% were observed in extrapulmonary TB cases (n = 86). Furthermore, significantly reduced MPT64+PstS1 concentrations (p < 0.001) were noticed in patients on therapy by RT-I-PCR as compared with untreated patients. Conclusion: Our RT-I-PCR assay revealed high sensitivity especially for the rapid diagnosis of smear-negative pulmonary TB and paucibacillary extrapulmonary TB samples, which could also monitor the dynamics of disease in patients on therapy.

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

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