Synthetic Peptides are Better Than Native Antigens for Development of ELISA Assay for Diagnosis of Tuberculosis

  • Arti R. Mishra
  • Vinita R. Hutke
  • Ashish R. Satav
  • Syed A. Ali
  • Hatim F. Daginawala
  • Lokendra R. Singh
  • Rajpal S. Kashyap


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.


Mycobacterium tuberculosis Tuberculous meningitis Serodiagnosis B-cell epitopes Peptide ELISA 



Tuberculous meningitis




Pulmonary tuberculosis


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


Mycobacterium tuberculosis


Meditation Addiction Health AIDS Nutrition


Non-tuberculous meningitis


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


Cerebrospinal fluid


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.

Informed Consents

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.

Supplementary material

10989_2016_9556_MOESM1_ESM.doc (490 kb)
Supplementary material 1 (DOC 490 kb)
10989_2016_9556_MOESM2_ESM.docx (34 kb)
Supplementary material 2 (DOCX 33 kb)
10989_2016_9556_MOESM3_ESM.docx (79 kb)
Supplementary material 3 (DOCX 79 kb)


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Arti R. Mishra
    • 1
  • Vinita R. Hutke
    • 1
  • Ashish R. Satav
    • 2
  • Syed A. Ali
    • 1
  • Hatim F. Daginawala
    • 1
  • Lokendra R. Singh
    • 1
  • Rajpal S. Kashyap
    • 1
  1. 1.Biochemistry Research LaboratoryCentral India Institute of Medical SciencesNagpurIndia
  2. 2.Meditations, AIDS, Health, Addiction & Nutrition (MAHAN) TrustC/O Mahatma Gandhi Tribal HospitalAmaravatiIndia

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