Title of article :
Expression and Purification of Recombinant Mycobacterium Tuberculosis (TB) Antigens, ESAT-6, CFP-10 and ESAT- 6/CFP-10 and Their Diagnosis Potential for Detection of TB Patients
Author/Authors :
Hemmati, M Department of Biochemistry - Recombinant Protein Laboratory - Medical School - Shiraz University of Medical Sciences, Shiraz , Seghatoleslam, A Department of Biochemistry - Recombinant Protein Laboratory - Medical School - Shiraz University of Medical Sciences, Shiraz , Rasti, M Department of Biochemistry - Recombinant Protein Laboratory - Medical School - Shiraz University of Medical Sciences, Shiraz , Ebadat, S Department of Biochemistry - Recombinant Protein Laboratory - Medical School - Shiraz University of Medical Sciences, Shiraz , Mosavari, N Department of PPD Tuberculin - Razi Vaccine and Serum Research Institute, Tehran , Habibagahi, M Department of Immunology - Medical School - Shiraz University of Medical Sciences, Shiraz , Taheri, M Department of Paramedical Sciences - Shiraz University of Medical Sciences, Shiraz , Sardarian, AR Dental School - Shiraz University of Medical Sciences, Shiraz , Mostafavi-Pour, Z Department of Biomedical Sciences - Shiraz University of Medical Sciences, Shiraz
Abstract :
Background: One of the most widely used methods to detect tuberculosis (TB) infection is the tuberculin skin
test (TST). The completion of Mycobacterium tuberculosis (M. tuberculosis) genome sequence has led to identification
of several antigens that can be utilized for accurate diagnosis and control of TB. The aim of this study
was to purify the recombinant M. tuberculosis antigens for the evaluation of their potential in TB diagnosis.
Methods: The recombinant secretory antigens, ESAT-6, CFP-10 and ESAT-6/CFP-10 were produced by PCR
and cloning methods. To investigate antigen specific responses of these recombinant antigens in detection of
TB, ex vivo enzyme linked immunospot (ELISPOT) test in 30 clinically diagnosed TB patients was evaluated.
Results: The selected M. tuberculosis antigens were cloned, expressed and purified in Escherichia coli (BL21).
ELISPOT assay for detection of TB showed the sensitivity of 93, 90 and 100% for recombinant ESAT-6, CFP-10
and ESAT-6/CFP-10 proteins respectively, which is significantly higher than conventional TST.
Conclusion: The recombinant antigens of ESAT-6, CFP-10 and ESAT-6/CFP-10 can be used as an accurate
means of detecting TB in Iran.
Keywords :
Mycobacterium tuberculosis , ELISPOT , CFP-10 , ESAT-6 , Tuberculosis , Diagnosis
