Diagnosis of Tuberculosis by Nanoparticle-Based Immuno-PCR Assay Based on Mycobacterial MPT64 and CFP-10 Detection

Abstract

Aim: To improve the diagnostic accuracy of immuno-PCR (I-PCR) in tuberculosis (TB) patients by using functionalized gold nanoparticles (AuNPs) coupled with detection antibodies and oligonucleotides, and magnetic beads (MBs) conjugated with capture antibodies in the liquid phase. Materials & methods: MB-coupled AuNP-based I-PCR (MB-AuNP-I-PCR) assay was designed to detect a cocktail of Mycobacterium tuberculosis MPT64 and CFP-10 proteins in bodily fluids of TB patients. Results: The sensitivities of 89.3 (n = 94) and 78.1% (n = 73) were observed in pulmonary TB and extrapulmonary TB patients, respectively, with specificities of 97.9–98.3%. Notably, the sensitivities attained by MB-AuNP-I-PCR in smear-negative pulmonary TB and extrapulmonary TB patients were significantly higher (p < 0.05–0.001) than Magneto-ELISA and GeneXpert assay. Conclusion: The improved technology, as well as enhanced diagnostic accuracy of MB-AuNP-I-PCR, may lead to development of an attractive diagnostic kit.

Keywords::

• diagnosis
• functionalized gold nanoparticles
• GeneXpert
• gold nanoparticles
• M. tuberculosis
• magnetic bead-coupled gold nanoparticle-based immuno-PCR
• magnetic beads
• magneto-ELISA
• MPT64+CFP-10
• tuberculosis

Supplementary data

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Financial & competing interests disclosure

Financial support provided by the Indian Council of Medical Research, Government of India, New Delhi (5/8/5/38/2014-ECD-I) and the Department of Biotechnology, Government of India, New Delhi (DBT; BT/PR8054/MED/29/710/2013) is acknowledged. The financial support obtained from JNU, New Delhi (JNU-UPE-II, ID-161) and JNU-DST-PURSE-II is also acknowledged. PK Mehta acknowledges the Council of Scientific and Industrial Research, New Delhi for awarding the Emeritus Scientist Fellowship, while B Dahiya acknowledges the DBT for providing the Junior/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

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.

Additional information

Funding

Financial support provided by the Indian Council of Medical Research, Government of India, New Delhi (5/8/5/38/2014-ECD-I) and the Department of Biotechnology, Government of India, New Delhi (DBT; BT/PR8054/MED/29/710/2013) is acknowledged. The financial support obtained from JNU, New Delhi (JNU-UPE-II, ID-161) and JNU-DST-PURSE-II is also acknowledged. PK Mehta acknowledges the Council of Scientific and Industrial Research, New Delhi for awarding the Emeritus Scientist Fellowship, while B Dahiya acknowledges the DBT for providing the Junior/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.