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)
IMPROVEMENT AND OPTIMIZATION
OF ANTIGENIC COMPOSITION FOR SERODIAGNOSIS OF
TUBERCULOSIS
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: saa0205@ukr.net
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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PDF (print version)
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.
Download
PDF (online version)
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: http://jvmbbs.kharkov.ua/archive/2016/volume2/issue4/oJVMBBS_2016024_011-015.pdf
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
References:
Banerjee, S., Gupta, S., Shende, N., Kumar, S.
and Harinath, B. C. (2003) ‘Serodiagnosis
of tuberculosis using two ELISA systems’, Indian Journal of Clinical
Biochemistry, 18(2), pp. 48–53. http://dx.doi.org/10.1007/bf02867367
Chambers, M. A., Whelan, A. O., Spallek, R.,
Singh, M., Coddeville, B., Guerardel,
Y. and Elass, E. (2010) ‘Non-acylated Mycobacterium bovis
glycoprotein MPB83 binds to TLR1/2
and stimulates production of matrix metalloproteinase 9’, Biochemical
and Biophysical Research Communications, 400(3), pp. 403–408. http://dx.doi.org/10.1016/j.bbrc.2010.08.085
Maia, R. P., Ask, B., Madsen, P., Pedersen, J. and Labouriau,
R. (2014) ‘Genetic determination of mortality rate in Danish dairy cows:
A multivariate competing risk analysis based on the number of survived
lactations’, Journal of Dairy Science, 97(3), pp. 1753–1761.
http://dx.doi.org/10.3168/jds.2013-6959
Manca,
C., Lyashchenko, K., Wiker,
H. G., Usai, D., Colangeli,
R. and Gennaro, M. L. (1997) ‘Molecular
cloning, purification, and serological characterization of MPT63, a novel antigen secreted by Mycobacterium
tuberculosis’, Infection and Immunity, 65(1), pp. 16–23.
Available at: http://iai.asm.org/content/65/1/16
Mencarini,
J., Cresci, C., Simonetti,
M. T., Truppa, C., Camiciottoli,
G., Frilli, M. L., Rogasi,
P. G., Veloci, S., Pistolesi,
M., Rossolini, G. M., Bartoloni,
A. and Bartalesi, F. (2016) ‘Non-tuberculous mycobacteria: Epidemiological pattern in a
reference laboratory and risk factors associated with pulmonary disease’,
Epidemiology and Infection, 145(03), pp. 515–522. http://dx.doi.org/10.1017/s0950268816002521
Miller, R. S., Farnsworth, M. L. and Malmberg,
J. L. (2013) ‘Diseases at the livestock-wildlife interface: Status,
challenges, and opportunities in the United States’, Preventive
Veterinary Medicine, 110(2), pp. 119–132. http://dx.doi.org/10.1016/j.prevetmed.2012.11.021
Redchuk,
T. A., Korotkevich, N. V., Kaberniuk,
A. A., Oliinyk, E. S., Labyntsev,
A. Y., Romaniuk, S. I., Kolibo,
D. V., Busol, V. A. and Komisarenko,
S. V. (2010) ‘Statistical analysis of the distribution of the antibody
levels to Mycobacterium bovis antigenes for bovine tuberculosis diagnostics’, Cytology
and Genetics, 44(5), pp. 280–285. http://dx.doi.org/10.3103/s009545271005004x
Siromolot,
A. A., Oliinyk, O. S., Kolibo,
D. V. and Komisarenko, S. V. (2016)
‘Mycobacterium tuberculosis antigens MPT63 and MPT83 increase phagocytic activity of murine peritoneal
macrophages’, The Ukrainian Biochemical Journal, 88(5), pp.
62–70. http://dx.doi.org/10.15407/ubj88.05.062
Wadhwa,
A., Johonson, R. E., Eda,
K., Waters, W., Palmer, M. V., Bannantine, J. P. and Eda, S. (2014) ‘Evaluation of ethanol vortex ELISA
for detection of bovine tuberculosis in cattle and deer’, BMC
Veterinary Research, 10(1), p. 147. http://dx.doi.org/10.1186/1746-6148-10-147
Waters, W. R., Buddle, B. M., Vordermeier, H. M., Gormley, E.,
Palmer, M. V., Thacker, T. C., Bannantine, J. P., Stabel, J. R., Linscott, R.,
Martel, E., Milian, F., Foshaug,
W. and Lawrence, J. C. (2011) ‘Development and evaluation of an
enzyme-linked immunosorbent assay for use in the
detection of bovine tuberculosis in cattle’, Clinical and Vaccine
Immunology, 18(11), pp. 1882–1888. http://dx.doi.org/10.1128/cvi.05343-11
Wiker,
H. G. (2009) ‘MPB70 and MPB83
— major antigens of Mycobacterium bovis’,
Scandinavian Journal of Immunology, 69(6), pp. 492–499. http://dx.doi.org/10.1111/j.1365-3083.2009.02256.x
Wiker,
H. G., Lyashchenko, K. P., Aksoy,
M. A., Lightbody, K. A., Pollock, J. M., Komissarenko, S. V., Bobrovnik,
S. O., Kolesnikova, I. N., Mykhalsky,
L. O., Gennaro, M. L. and Harboe,
M. (1998) ‘Immunochemical characterization of the MPB70/80
and MPB83 proteins of Mycobacterium bovis’, Infection and Immunity, 66(4), pp.
1445–1452. Available at: http://iai.asm.org/content/66/4/1445
