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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
Article
  • 96 Downloads

Abstract

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.

Keywords

Mycobacterium tuberculosis Tuberculous meningitis Serodiagnosis B-cell epitopes Peptide ELISA 

Abbrevations

TBM

Tuberculous meningitis

TB

Tuberculosis

PTB

Pulmonary tuberculosis

EPTB

Extra pulmonary tuberculosis

ELISA

Enzyme-linked immunosorbent assay

PCR

Polymerase chain reaction

Ag85B

Antigen 85 B

45 kDa

45 kilo Dalton

HSP 16

Heat Shock Protein 16

CFP-10

10 kDa culture filtrate protein

ESAT-6

6 kDa Early secretory antigenic target

CIIMS

Central India Institute of Medical Sciences

MTB

Mycobacterium tuberculosis

MAHAN

Meditation Addiction Health AIDS Nutrition

NTBM

Non-tuberculous meningitis

NPTB

Non pulmonary tuberculosis

WHO

World Health Organization

MIF

Molecular Immunology Foundation-Bioinformatics software

Bcepred

Prediction of linear B-cell epitopes—Bioinformatics software

ABCpred

Artificial neural network based B-cell epitope prediction server

NCBI

National Center for Biotechnology Information

BLAST

Basic local alignment search tool

CSF

Cerebrospinal fluid

ATT

Anti tuberculosis treatment

QFT

QuantiFERON-TB Gold test

TST

Tuberculin skin test

Notes

Acknowledgments

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’.

Funding

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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