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
نویسندگان
1 Department of Biochemistry, Recombinant Protein Laboratory, Medical School, Shiraz University of Medical Sciences, Shiraz,
2 Department of Biochemistry, Recombinant Protein Laboratory, Medical School, Shiraz University of Medical Sciences, Shiraz,
3 Department of PPD Tuberculin, Razi Vaccine and Serum Research Institute, Tehran,
4 Department of Immunology, Medical School,
5 Department of Paramedical Sciences,
6 Department of Biochemistry, Recombinant Protein Laboratory, Medical School, Shiraz University of Medical Sciences, Shiraz,
7 Department of Biochemistry, Recombinant Protein Laboratory, Medical School, Shiraz University of Medical Sciences, Shiraz,
8 Department of Biochemistry, Recombinant Protein Laboratory, Medical School, Shiraz University of Medical Sciences, Shiraz,
9 Dental School,
doi
چکیده
Background: One of the most widely used methods to detect tuberculosis (TB) infection is the tuberculin skintest (TST). The completion of Mycobacterium tuberculosis (M. tuberculosis) genome sequence has led to identificationof several antigens that can be utilized for accurate diagnosis and control of TB. The aim of this studywas 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 PCRand cloning methods. To investigate antigen specific responses of these recombinant antigens in detection ofTB, 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-10and 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 accuratemeans of detecting TB in Iran.
