Development of a New Indirect ELISA Method for Detection of Anti-Tuberculosis Antibodies in Human Serum
Abstract
Background: Tuberculosis is a crucial health problem. Establishing a rapid, reliable and still inexpensive diagnostic method for tuberculosis seems to be substantial in developing countries where TB has very high incidence rate .
Methods: An Indirect Enzyme-linked immunosorbent Assay (ELISA) was established to detect serum antibodies against Mycobacterium tuberculosis. Three kinds of antigens were used to prepare the solid phase for antibody as- say including: purified protein derivative (PPD), M. tuberculosis Bacilli, and Mycobacterium bovis Bacillus Calmette Guerin (BCG). Sera of two main following groups were investigated in this study: sera samples from smear- positive, culturepositive and Tuberculin Skin Test-positive TB patients and sera samples from smear-negative, culture negative and TST-negative healthy individuals.
Results: Among the antigens used, BCG produced higher sensitivity and specificity in the assay. With PPD as the solid phase, higher sensitivity but low er specificity was observed in comparison with BCG. Both, low response and noise (non-specific binding) were observed with TB bacilli as the solid phase in the assay.
Conclusion: Using BCG solid phase system in this method resulted in higher sensitivity in comparison to single antigen solid phase systems. In addition, we were able to circumvent the problem of non-specific bindings in more popular multi-antigenic solid systems such as PPD. By using this new indi- rect ELISA, a rapid, reliable and still inexpensive diagnosis of tuberculosis might be possible. Although, further investigations are required to confirm our result.
WHO (World Health Organization).Global tuberculosis control:WHO Re port Geneva 2010.
Dye C, Scheele S, Dolin P, et al. Con sensus statement. Global burden of tuberculosis: estimated incidence, pre valence, and mortality by country. W- HO Global Surveillance and Mon- itoring Project. JAMA 1999; 282 (7):677-86.
Behr MA, Warren SA, Salamon H, et al. Transmission of Mycobacterium tu berculosis from patients smearnegati ve for acid-fast bacilli. Lancet 199- 9;353 (9151): 444-9.
Brown S, Petrof S. The Clinical Value of complement fixation in pulmonary tuberculosis based on a study of 540 cases. Am Rev Tuberc 1918; 2: 525-40.
Glatman-Freedman A. Advances in antibody-mediated immunity against Mycobacterium tuberculosis: implica tions for a novel vaccine strategy. FE MS Immunol Med Microbiol 2003; 39 (1): 9-16.
Glatman-Freedman A, Martin JM, Riska PF, et al. Monoclonal antibo- dies to surface antigens of Mycobac- terium tuberculosis and their use in a modified enzyme-linked immuno- sorbent spot assay for detection ofmycobacteria. J Clin Microbiol 1996;34 (11): 2795-802.
Glatman-Freedman A, Casadevall A, Dai Z, et al. Antigenic evidence of pre valence and diversity of Mycobacterium tuberculosis arabinomannan. J Clin Microbiol 2004; 42 (7): 3225-31.
Chan ED, Heifets L, Iseman MD. Immunologic diagnosis of tuberculosis: a review. Tuber Lung Dis 2000; 80 (3):131-40.
Garg SK, Tiwari RP, Tiwari D, et al. Diagnosis of tuberculosis: available tec hnologies, limitations, and possibilities. J Clin Lab Anal 2003; 17 (5): 155-63.
Pereira Arias-Bouda LM, Nguyen LN, Ho LM, et al. Development of antigen detection assay for diagnosis of tuberculosis using sputum samples.J Clin Microbiol 2000; 38 (6): 2278-83.
Pereira Arias-Bouda LM, Kuijper S,van Deutekom H, et al. Enzyme-linked immunosorbent assays using immune complexes for the diagnosis of tubercu-losis. J Immunolo methods 2003; 283(1-2): 115-24.
Seeley JH, Vandemark P, Lee J.,1997.Medical microbiology and immunology. In: A laboratory manual of microbiology. W. H. Freeman and company, New York.James SH, Dumois JA 3rd, Messina AF, et al. Healthcare worker know- ledge of measurement and docu- mentation of tuberculin skin test reac- tion. Infect Control Hosp Epidemiol 2009; 30 (12): 1230-2.
Pottumarthy S, Morris AJ, Harrison AC, et al. Evaluation of the tuberculin gamma interferon assay: potential to replace the Mantoux skin test. J Clin Microbiol. 1999; 37 (10): 3229-32.
Sohn H, Minion J, Albert H, et al. TB diagnostic tests: how do we figure out their costs? Expert Rev Anti Infect Ther 2009; 7 (6): 723-33.
Dowdy DW, Steingart KR, Pai M. Serological testing versus other strat- egies for diagnosis of active tubercu losis in India: a cost-effectiveness ana lysis. PLoS Med 2011 ; 8 (8): e100 1074.
Farajollahi MM, Cook DB, Hamzeh-lou S, et al. Reduction of non-specific binding in immunoassays requiring long incubations. Scand J Clin Lab Invest 2012; 72 (7): 531-9.
Barka NE, Agopian MS, Peter JB. False-positive IgM antibodies to Bor- relia burgdorferi in indirect ELISA as a result of IgM rheumatoid factor. J Infect Dis 1990; 161 (6): 1312.
Ramos-Levi AM, Montanez MC, Ortega I, et al. A case of biochemical assay discrepancy: Interference with measurement of thyroid-stimulating hormone due to rheumatoid factor.Endocrinol Nutr 2012 [In press].
Bartels EM, Ribel-Madsen S. Cyto- kine measurements and possible in- terference from heterophilic antibo- diesProblems and solutions experie nced with rheumatoid factor. Methods 2013 [In press].
Churchman SM, Geiler J, Parmar R,et al. Multiplexing immunoassays for cytokine detection in the serum of pa- tients with rheumatoid arthritis: lack of sensitivity and interference by rheumatoid factor. Clin Exp Rheumatol 2012; 30 (4): 534-42.
al-Hajjaj MS, Gad-el-Rab MO, al Orainey IO, et al. Improved sensitivity for detection of tuberculosis cases by a modified Anda-TB ELISA test. Tuber Lung Dis 1999; 79 (3): 181-5.
Houghton RL, Lodes MJ, Dillon DC, et al. Use of multiepitope polyproteins in serodiagnosis of active tuberculosis.Clin Diagn Lab Immunol 2002; 9 (4):883-91.
Panda D, Lahiri A, Bhattacharyya I, et al. Humoral immune responses in different clinical forms of tuberculosis. J Indian Med Assoc 2001; 99 (8): 424,426-7, 440.
Simonney N, Molina JM, Molimard M, et al. Comparison of A60 and three glycolipid antigens in an ELISA test for tuberculosis. Clin Microbiol Infect 1996; 2 (3): 214-22.
Whelan C, Shuralev E, Kwok HF, et al. Use of a multiplex enzyme-linked immunosorbent assay to detect a sub- population of Mycobacterium bovisinfected animals deemed negative or inconclusive by the single intradermal comparative tuberculin skin test. J Vet Diagn Invest 2011; 23 (3): 499-503.
Sumi S, Radhakrishnan VV. Diagnostic significance of humoral immune responses to recombinant antigens of Mycobacterium tuberculosis in pa- tients with pleural tuberculosis. J Clin Lab Anal 2010; 24 (5): 283-8.
da Silva EB, Silva BD, Leon JR, et al.Using BCG, MPT-51 and Ag85 as antigens in an indirect ELISA for the diagnosis of bovine tuberculosis. Vet J 2011; 187 (2): 276-8.
Gaborick CM, Salman MD, Ellis RP,et al. Evaluation of a five-antigen ELI SA for diagnosis of tuberculosis in cattle and Cervidae. J Am Vet Med Assoc 1996; 209 (5): 962-6.
Files | ||
Issue | Vol 1 No 3-4 (2012) | |
Section | Original Articles | |
Keywords | ||
BCG Vaccine Tuberculosis Enzyme-Linked Immunosorbent Assay |
Rights and permissions | |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |