Issue 4

Journal for Veterinary Medicine, Biotechnology and Biosafety

Volume 2, Issue 4, December 2016, Pages 11–15

ISSN 2411-3174 (print version) ISSN 2411-0388 (online version)


Siromolot A. A. 1, 2 , Oliinyk O. S. 2 , Kolibo D. V. 1, 2 , Gerilovych A. P. 3

1 Educational and Scientific Centre, Institute of Biology and Medicine, Taras Shevchenko National University of Kyiv, Ukraine, e-mail:

2 Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, Kyiv, Ukraine

3 National Scientific Center ‘Institute of Experimental and Clinical Veterinary Medicine’, Kharkiv, Ukraine

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Citation for print version: Siromolot, A. A., Oliinyk, O. S., Kolibo, D. V. and Gerilovych, A. P. (2016) ‘Improvement and optimization of antigenic composition for serodiagnosis of Tuberculosis’, Journal for Veterinary Medicine, Biotechnology and Biosafety, 2(4), pp. 11–15.

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Citation for online version: Siromolot, A. A., Oliinyk, O. S., Kolibo, D. V. and Gerilovych, A. P. (2016) ‘Improvement and optimization of antigenic composition for serodiagnosis of Tuberculosis’, Journal for Veterinary Medicine, Biotechnology and Biosafety. [Online] 2(4), pp. 11–15. Available at:

Summary. There is a pressing of needs for improvement existing methods of tuberculosis (TB) diagnosis and screening. New methods should be characterized by high specificity, sensitivity, reliability of results; easiness of implementation; precision interpretation of the results. Regarding the TB, the ELISA test-systems have many advantages over traditional methods of disease diagnostics. However, the most difficult objective of creating such test-systems is the selection of optimal antigenic substance which would characterized by the high antigenicity, and high specificity from the other hand. The aim of the research was to improve previously created antigenic composition using full-size highly immunogenic proteins of Mycobacterium tuberculosis MPT63 and MPT83. The genetic construction pET28a-MPT83(full)-MPT63 was obtain, with further isolation and purification of target protein and compared to its predecessor which based on FLD of MPT83(115–220 aa). Both fusion proteins were tested to the culture medium of hybridomas, which were obtained from mice immunized with a mixture of antigens from mycobacteria, including cell membrane, associated protein MPT83 closest homologue — MPT70. In addition, sera samples from infected M. bovis and healthy cattle were tested on several variants of Mycobacterium fusion proteins. These data suggest the need of N-terminal amino acids for protein folding for better antibody recognition. The new antigenic substance gives the best results for serology-based diagnostics of tuberculosis. Using of specific monoclonal antibodies to new obtained antigen as a positive control for the test system can reduce the cost of it and avoid using genuine infected cattle or human serum among ELISA kit reagents.

Keywords: Mycobacterium tuberculosis, Mycobacterium bovis, antigenic substance, MPT63, MPT83, chimeric protein, serology-based diagnostics, tuberculosis


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