International E-publication: Publish Projects, Dissertation, Theses, Books, Souvenir, Conference Proceeding with ISBN.  International E-Bulletin: Information/News regarding: Academics and Research

Rapid Immunodiagnosis of Tuberculosis by using Polymerase Chain Reaction and in-house developed Enzyme linked Immunosorbent Assay

Author Affiliations

  • 1 Department of Microbiology, Karpagam University, Coimbatore - 641021, Tamil Nadu, INDIA

Int. Res. J. Biological Sci., Volume 2, Issue (8), Pages 42-47, August,10 (2013)


Tuberculosis (TB) remains the second leading cause of death among all infected individuals and is responsible for 2 million deaths annually. The definitive diagnosis of TB is by demonstrating acid-fast bacilli either by direct smear or culture. Since these methods are insensitive (smear technique) and time consuming (culture technique), additional immunodiagnostic methods such as antigen or antibody demonstration has become necessary. The main purpose of this study is to develop a diagnostic tool which can yield results in short span of time with much higher sensitivity rate. In this study, 200 serum samples were used for the development of ELISA technique. Out of 200 samples, 149 gave positive results and the sensitivity of the developed test appeared to be 88.7%.


  1. Mendiratta D.K., Narang P. and Narang R., Face to face with non tuberculosis mycobacteria at Sevagram, Rev. J. MGIMS, 14, 16-21 (2009)
  2. Weir R.E., Black G.F., Nazareth B., Floyd S., Stenson S., Stanley C., Branson K., Sichali L., Chaguluka S.D., Donovan L., Crampin A.C., Fine P.E. and Dockrell H.M., The influence of previous exposure to environmental mycobacteria on the interferon-gamma response to Bacille Calmette- Guerin vaccination in southern England and northern Malawi, Clin. Exp. Immunol.,146, 390-399 (2006)
  3. Lyashchenko K., Colangeli R., Houde M., Jahdali H.A., Menzies D. and Gennaro M.L., Heterogeneous antibody responses in tuberculosis, Infect. Immun.,66, 3936-3940 (1998)
  4. Houghton R.L., Lodes M.J., Dillon D.C., Reynolds L.D., Day C.H. and McNeill P.D., Use of multiepitope polyproteins in serodiagnosis of active tuberculosis, Clin. Diag. Lab. Immunol.,9, 883-891 (2002)
  5. Mukherjee S., Daifalla N., Zhang Y., Douglass J., Brooks L. and Vedvick T., Potential serological use of a recombinant protein that is a replica of a Mycobacterium tuberculosis protein found in the urine of infected mice, Clin. Diagn. Lab. Immunol.,11, 280-286 (2004)
  6. Deshpande P.S., Kashyap R.S., Ramteke S.S., Nagdev K.J., Purohit H.J., Taori G.M. and Daginawala H.F., Evaluation of the IS6110 PCR assay for the rapid diagnosis of tuberculous meningitis, Cerebrospinal Fluid Res.,:10 2007)
  7. Lowry O.H., Rosebrough N.J., Farr A.L. and Randall R.J., Protein measurement with the Folin-phenol reagent, J. Biol. Chem.,193, 265275 (1951)
  8. Nath S.S., Krishnan N., Angayarkanni J. and Palaniswamy M., Detection of mycobacterial antibodies in serum samples using BCG vaccine based ELISA, Int. J. Pharma. Pharma. Sci.,4 (Suppl 1), 318-321 (2012)
  9. Nath S.S., Krishnan N. and Palaniswamy M., Vaccine turned to diagnostic tool, Asian J. Pharma. Clin. Res.,5 (Suppl 2), 54-56 (2012)
  10. Fujiwara N., Distribution of antigenic glycolipids among Mycobacterium tuberculosis strains and their contribution to virulence, Kekkaku., 72, 193205 (1997)
  11. Wadee A.A., Boting L. and Reddy S.G., Antigen capture assay for detection of a 43- kilodalton Mycobacterium tuberculosis antigen, J. Clin. Microbiol., 28(12), 2786-2791 (1990)
  12. Harinath B.C., Kumar S., Saha-Roy S., Hirudkar S., Upadhye V. and Shende N., A cocktail of affinity-purified antibodies reactive with diagnostically useful mycobacterial antigens ES-31, ES-43 and EST-6 for detecting the presence of Mycobacterium tuberculosis, Diagn. Microbiol. Infect. Dis., 55, 65-68 (2006)
  13. Gutierrez E.A., Amezcua-Chavarria M.E. and Paten-Sanclezy S., Enzyme linked immunosorbant assay (ELISA) with mycobacterial crude antigens for the sero-epidemiological diagnosis of active tuberculosis, Int. J. Lepr.,64, 417-427 (1996)
  14. Ridell M., Waller S.G., Minnikin D.E., Bolten R.C. and Magnusson M., A comparative serological study of antigenic glycolipids from Mycobacterium tuberculosis, Tubercle Lung Dis.,73, 101-105 (1992)
  15. Ohman R. and Ridell M., Purification and characterization of isocitrate dehydrogenase and malate dehydrogenase from M. tuberculosis and evaluation of their potential as suitable antigens for the diagnosis of tuberculosis, Tubercle Lung Dis.,77, 454-461 (1996)
  16. van Pinxteren L.A., Ravn P., Agger E.M., Pollock J. and Andersen P., Diagnosis of tuberculosis based on the two specific antigens ESAT-6 and CFP10, Clin. Diagn. Lab. Immunol.,7, 155-160 (2000)
  17. Sada E., Ruiz-Palacios G.M., Lopez-Vidal Y. and Ponce de Leon S., Detection of mycobacterial antigens in cerebrospinal fluid of patients with tuberculous meningitis by enzyme-linked immunosorbent assay, Lancet,17, 651652 (1983)
  18. Charpin D., Herbault H., Gevaudan M.J., Saadjian M., de Mico P., Arnaud A., Vervloet D. and Charpin J., Value of ELISA using A60 antigen in the diagnosis of active pulmonary tuberculosis, Am. Rev. Respir. Dis.,142, 380384 (1990)
  19. Cocito C.G., Properties of the mycobacterial antigen complex A60 and its applications to the diagnosis and prognosis of tuberculosis, Chest, 100, 16781693 (1991)
  20. Turneer M.E., Van Nerom J., Nyabenda J., Waelbroeck A., Duvivier A. and Toppet M., Determination of humoral immunoglobulins M and G directed against mycobacterial antigen 60 failed to diagnose primary tuberculosis and mycobacterial adenitis in children, Am. J. Respir. Crit. Care Med.,150, 15081512 (1994)
  21. Gupta S., Kumari S., Banwalikar J.N. and Gupta S.K., Diagnostic utility of the estimation of mycobacterial antigen A60 specific immunoglobulins IgM, IgA, and IgG in the sera of cases of adult human tuberculosis, Tuber. Lung Dis.,76, 418424 (1995)
  22. Adjei A.A., Armah H., Achaw O., Adiku T. and Hesse I.F.A., Evaluation of a rapid serological chromatographic immunoassay for the diagnosis of pulmonary tuberculosis in Accara, Ghana Jjid.,56, 161164 (2003)
  23. Stavri H., Moldovan O., Mihaltan F., Banica D. and Doyle R.J., Rapid dot sputum and serum assay in pulmonarym tuberculosis, J. Microbiol. Meth., 52, 285296 (2003)