Synthetic Peptides are Better Than Native Antigens for Development of ELISA Assay for Diagnosis of Tuberculosis
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Immunodiagnosis of both pulmonary tuberculosis (PTB) and extrapulmonary tuberculosis has remained challenging. In the present work, in-house developed synthetic peptide based antibody detection assay was assessed and validated with antigen based assay for effective diagnosis of tuberculosis (TB). The study population included both tuberculous meningitis (TBM) (n = 60) admitted to Neurology IPD wards of our Institute hospital and PTB cases (n = 57) recruited from high TB endemic zones. Peptides of five highly immunogenic Mycobacterium tuberculosis (MTB) proteins (Ag85B, 45 kDa, HSP-16, CFP-10 and ESAT-6) were designed and synthesized. The designed peptides were evaluated in samples of both TBM and PTB cases, respectively, using in-house developed antibody detection method. The developed tests were further compared and validated with MTB native proteins based antibody detection ELISA. Sensitivity and specificity of peptide assay were significantly higher or almost similar (p < 0.05) in TBM and PTB as compared to native proteins based ELISA. Among all peptides, diagnostic reliability of Ag 85B peptide A1 was higher for both forms of TB. Peptide-based antibody assay is cost effective, simple and may be interchangeable with conventional Antigen based ELISA assays for effective diagnosis of TB in the developing world.
KeywordsMycobacterium tuberculosis Tuberculous meningitis Serodiagnosis B-cell epitopes Peptide ELISA
Extra pulmonary tuberculosis
Enzyme-linked immunosorbent assay
Polymerase chain reaction
Antigen 85 B
- 45 kDa
45 kilo Dalton
- HSP 16
Heat Shock Protein 16
10 kDa culture filtrate protein
6 kDa Early secretory antigenic target
Central India Institute of Medical Sciences
Meditation Addiction Health AIDS Nutrition
Non pulmonary tuberculosis
World Health Organization
Molecular Immunology Foundation-Bioinformatics software
Prediction of linear B-cell epitopes—Bioinformatics software
Artificial neural network based B-cell epitope prediction server
National Center for Biotechnology Information
Basic local alignment search tool
Anti tuberculosis treatment
QuantiFERON-TB Gold test
Tuberculin skin test
The authors would like to thank Dr. Shradha Bhullar for assistance with designing of peptides, and Dr. Ali Abbas Husain for comments that greatly improved the manuscript. We would likewise wish to acknowledge the assistance of Colorado State University, USA for providing us with tuberculosis research material (Contract No. 1-A1-40091 entitled ‘Tuberculosis Research Material and Vaccine Testing’.
This study was funded by Central India Institute of Medical Sciences, Nagpur as a part of its in house study.
Compliance with Ethical Standards
Conflicts of Interest
The authors affirm that this article content has no conflict of interest.
The study was approved by the Institutional Ethics Committee. Written consents were obtained from each participant and oral explanation about the research study was presented to all participants.
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