Proteomic analysis of purified protein derivative of Mycobacterium tuberculosis

Clinical Proteomics

Jayasuryan Narayana³  HC Harsha⁷  Akhilesh Pandey⁸  Aditi Chatterjee¹  Vithal P Myneedu⁶  Anjali Ganjiwale³  Vinuth N Puttamallesh⁷  Jyoti Sharma¹  Anil K Madugundu⁷  Gajanan J Sathe¹  Raja Sekhar Nirujogi²  Satish Kumar⁴  Renu Verma³  Thottethodi Subrahmanya Keshava Prasad⁵ 

[1] Manipal University, Madhav Nagar, Manipal 576 104, India;Centre for Bioinformatics, School of Life Sciences, Pondicherry University, Puducherry 605 014, India;Microtest Innovations Pvt. Ltd., International Tech Park, Whitefield, Bangalore 560 066, India;Armed Forces Medical College, Pune 411 040, India;School of Biotechnology, KIIT University, Bhubaneswar, Odisha-751024, India;Department of Microbiology, LRS Institute of TB and RD, New Delhi 110 030, India;Institute of Bioinformatics, International Tech Park, Whitefield, Bangalore 560 066, India;Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA

关键词: Mantoux test;    LC-MS/MS;    Epitope;    Broad spectrum antibiotics;    Biomarker;   

Others  :  1026342
DOI  :  10.1186/1559-0275-10-8

 received in 2013-03-28, accepted in 2013-07-16,  发布年份 2013

【 摘 要 】

Background

Purified protein derivative (PPD) has been used for more than half a century as an antigen for the diagnosis of tuberculosis infection based on delayed type hypersensitivity. Although designated as “purified,” in reality, the composition of PPD is highly complex and remains ill-defined. In this report, high resolution mass spectrometry was applied to understand the complexity of its constituent components. A comparative proteomic analysis of various PPD preparations and their functional characterization is likely to help in short-listing the relevant antigens required to prepare a less complex and more potent reagent for diagnostic purposes.

Results

Proteomic analysis of Connaught Tuberculin 68 (PPD-CT68), a tuberculin preparation generated from M. tuberculosis, was carried out in this study. PPD-CT68 is the protein component of a commercially available tuberculin preparation, Tubersol, which is used for tuberculin skin testing. Using a high resolution LTQ-Orbitrap Velos mass spectrometer, we identified 265 different proteins. The identified proteins were compared with those identified from PPD M. bovis, PPD M. avium and PPD-S2 from previous mass spectrometry-based studies. In all, 142 proteins were found to be shared between PPD-CT68 and PPD-S2 preparations. Out of the 354 proteins from M. tuberculosis–derived PPDs (i.e. proteins in either PPD-CT68 or PPD-S2), 37 proteins were found to be shared with M. avium PPD and 80 were shared with M. bovis PPD. Alignment of PPD-CT68 proteins with proteins encoded by 24 lung infecting bacteria revealed a number of similar proteins (206 bacterial proteins shared epitopes with 47 PPD-CT68 proteins), which could potentially be involved in causing cross-reactivity. The data have been deposited to the ProteomeXchange with identifier PXD000377.

Conclusions

Proteomic and bioinformatics analysis of different PPD preparations revealed commonly and differentially represented proteins. This information could help in delineating the relevant antigens represented in various PPDs, which could further lead to development of a lesser complex and better defined skin test antigen with a higher specificity and sensitivity.

【 授权许可】

   
2013 Prasad et al.; licensee BioMed Central Ltd.

【 图 表 】

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