THE SEARCH OF OPTIMAL COMBINATION OF ANTIGENS FOR SEROLOGICAL DIAGNOSTICS OF TUBERCULOSIS

Cover Page


Cite item

Full Text

Abstract

Abstract. The four chimeric recombinant antigens CBD-CFP10, CBD-ESAT6, ESAT6-CFP10 and CBD-P38 contained aminoacid sequences of full-size proteins ESAT6, CFP10 and matured protein P38 of M. tuberculosis, joined with aminoacid sequences of cellulose bind domain of endogluconase A (CBD) from Cellumonas fimi have been obtained by gene engineering methods. Recombinant proteins were purified by affine chromatography in column with Ni-NTA-sepharose 6В-CL and as PPDN-3 were used for detection of their antigenic activity in indirect ELISA for TB serological diagnostics. The sera from patients with lung tuberculosis (n = 321), from persons who had professional contacts with TB patients (n = 42), from healthy blood donors (n = 366) and from patients with lung diseases of non-TB etiology were tested. It was detected that there was positive correlation between antibodies level for all studied antigens compared by pair. It has been demonstrated that although antigens were different by antigenic and immunobiological characteristics they add each other in the content of antigenic diagnostics compositions. Thus, all these antigens can be used in the test kits for serological diagnostics of TB. Using of these antigens will allow to detect persons infected by TB and patients with active tuberculosis. 

About the authors

E. V. Vasilyeva

Saint Petersburg Pasteur Institute, Saint Petersburg

Author for correspondence.
Email: ev_vasilyeva@pasteurorg.ru

Junior Research Associate, Department of New Technologies, Laboratory of Biological Preparations

Mira str. 14, St. Petersburg, 197101, Russian Federation

 

Russian Federation

V. N. Verbov

Saint Petersburg Pasteur Institute, Saint Petersburg

Email: ev_vasilyeva@pasteurorg.ru
PhD (Chemistry), Chief, New Technologies Department Russian Federation

A. B. Beklemishev

Research Institute for Biochemistry of Siberian Branch of the Russian Academy of Medical Sciences, Novosibirsk

Email: ev_vasilyeva@pasteurorg.ru
PhD, MD (Biology), Professor, Chief, Laboratory of Genetic Engineering Russian Federation

A. L. Mamaev

Research Institute for Biochemistry of Siberian Branch of the Russian Academy of Medical Sciences, Novosibirsk

Email: ev_vasilyeva@pasteurorg.ru
Junior Research Associate Russian Federation

A. O. Tsyrul’nikov

Research Institute for Biochemistry of Siberian Branch of the Russian Academy of Medical Sciences, Novosibirsk

Email: ev_vasilyeva@pasteurorg.ru

Junior Research Associate 

Russian Federation

T. A. Stepanenko

City Multiprofile hospital No. 2, St. Petersburg

Email: ev_vasilyeva@pasteurorg.ru
PhD (Medicine), Head of the Department of Pulmonology No. 2 Russian Federation

V. B. Ivanovskii

City Tuberculosis Dispensary, St. Petersburg

Email: ev_vasilyeva@pasteurorg.ru
PhD (Medicine), Deputy Chief Physician on Medical Work Russian Federation

S. V. Lapin

Pavlov State Medical University, St. Petersburg

Email: ev_vasilyeva@pasteurorg.ru
PhD (Medicine), Chief, Laboratory of Autoimmune Diagnostics, Center for Molecular Medicine Russian Federation

Areg A. Totolian

Saint Petersburg Pasteur Institute, Saint Petersburg

Email: ev_vasilyeva@pasteurorg.ru
PhD, MD (Medicine), Corresponding Member of the RAMS, Deputy Director on Scientific Work Russian Federation

References

  1. Васильева Е.В., Вербов В.Н., Тотолян Арег А., Беклемишев А.Б., Мамаев А.Л., Цырульников А.О. Возможность использования различных антигенов M. tuberculosis в серодиагностике туберкулеза легких // Медицинский академический журнал. — 2012. — № 4 (12). — С. 24–26. Vasil`eva E.V., Verbov V.N., Totolyan Areg A., Beklemishev A.B., Mamaev A.L., Tsirul`nikov A.O. Vozmozhnost` ispol`zovaniya razlichnykh antigenov M. tuberculosis v serodiagnostike tuberkuleza legkikh [Possibilities of using of different M. tuberculosis antigens in serological diagnosis of lung tuberculosis]. Meditsinskiy akademicheskiy zhurnal — Medical Academic Journal, 2012, vol. 12, no. 4, pp. 24–26.
  2. Деркачева С.А., Ткаченко В.И., Беклемишев А.Б. Получение рекомбинантного антигена р38 M. tuberculosis и оценка возможности его применения в серодиагностике туберкулеза // Бюллетень СО РАМН. — 2007. — № 5 (127). — С. 62–68. Derkachiova S.A., Tkachenko V.I., Beklemishev A.B. Poluchenie rekombinantnogo antigena r38 M. tuberculosis i otsenka vozmozhnosti ego primeneniya v serodiagnostike tuberkulioza [Obtaining of recombinant antigen of M. tuberculosis and evaluation of possibilities its using in serological diagnostics of tuberculosis]. Byulleten` SO RAMN — Bulletin of the Siberian Branch of the Russian Academy of Medical Sciences, 2007, vol. 127, no. 5, pp. 62–68.
  3. Маниатис Т., Фрич Э., Сэмбрук Дж. Методы генетической инженерии. Молекулярное клонирование // М., 1984. — 480 с. Maniatis T., Frich E., Sembruk Dzh. Metody geneticheskoy inzhenerii. Molekulyarnoe klonirovanie [Methods of gene engineering. Molecular cloning]. Moscow, 1984. 480 p.
  4. Салина Т.Ю., Морозова Т.И. Иммунологические методы в дифференциальной диагностике // Туберкулез и болезни легких. — 2011. — № 11. — C. 50–53. Salina T.Yu., Morozova T.I. Immunologicheskie metody v differentsial`noy diagnostike [Immunological methods in differential diagnostics]. Tuberkulez i bolezni legkikh — Tuberculosis and Lung Diseases, 2011, no. 11, pp. 50–53.
  5. Туберкулез в Российской Федерации, 2010 г. Аналитический обзор статистических показателей, используемых в Российской Федерации. — М., 2011. — 280 с. Tuberkulez v Rossiyskoy Federatsii, 2010 g. Analiticheskiy obzor statisticheskikh pokazateley, ispol`zuemykh v Rossiyskoy Federatsii [Tuberculosis in the Russian Federation, 2010. Analytical review of statistical indices, used in the Russian Federation]. Moscow, 2011. 280 p.
  6. Chegou N.N., Hoek K.G., Kriel M., Warren R.M., Victor T.C., Walzl G. Tuberculosis assays: past, present and future. Expert Rev. Anti Infect. Ther., 2011, vol. 9, no. 4, pp. 457–469.
  7. Cole S.T., Brosch R., Parkhill J. Deciphering the biology of Mycobacterium Tuberculosis from the complete genome sequence. Nature, 1998, vol. 393, no. 6685, pp. 537–544.
  8. Davidow A., Kanaujia G.V., Shi L., Kaviar J., Guo X., Sung N., Kaplan G., Menzies D., Gennaro M.L. Antibody profiles characteristic of Mycobacterium tuberculosis infection state. Infect. Immun., 2005, vol. 73, no. 10, pp. 6846–6851.
  9. Ireton G.C., Greenwald R., Liang H., Esfandiari J., Lyashchenko K.P., Reed S.G. Identification of Mycobacterium tuberculosis antigens of high serodiagnostic value. Clin. Vaccine Immunol., 2010, vol. 17, no. 10, pp. 1539–1547.
  10. Hatfield G. W., Roth D.A. Optimizing scaleup yield for protein production: Computationally Optimized DNA Assembly [CODA] and Translation Engineering. Biotechnology Annual Review, 2007, vol. 13, pp. 27–42.
  11. He X.Y., Li J., Hao J., Chen H.B., Zhao Y.Z., Huang X.Y., He K., Xiao L., Ye L.P., Qu Y.M., Ge L.H. Assessment of five antigens from Mycobacterium tuberculosis for serodiagnosis of tuberculosis. Clin. Vaccine Immunol., 2011, vol. 18, no. 4, pp. 565–570.
  12. Manca C., Lyashchenko K., Wiker H.G., Usai D., Colangeli R., Gennaro M.L. Molecular cloning, purification, and serological characterization of MPT63, a novel antigen secreted by Mycobacterium tuberculosis. Infect. Immun., 1997, vol. 65, no. 1, pp. 16–23.
  13. Min F., Zhang Y., Huang R., Li W., Wu Y., Pan J., Zhao W., Liu X. Serum antibody responses to 10 Mycobacterium tuberculosis proteins, purified protein derivative, and old tuberculin in natural and experimental tuberculosis in rhesus monkeys. Clin. Vaccine Immunol., 2011, vol. 18, no. 12, pp. 2154–2160.
  14. Pinto L.M., Grenier J., Schumacher S.G., Denkinger C.M., Steingart K.R., Pai M. Immunodiagnosis of tuberculosis: state of the art. Med. Princ. Pract., 2012, vol. 21, no. 1, pp. 4–13.
  15. Randall L.L., Hardy S. J. S. Correlation of competence for export with lack of tertiary structure of the mature species: a study in vivo of maltose-binding protein in E. coli. Cell, 1986, vol. 46, pp. 921–928.
  16. Singh S., Singh J., Kumar S., Gopinath K., Balooni V., Singh N., Mani K. Poor performance of serological tests in the diagnosis of pulmonary tuberculosis: evidence from a contact tracing field study. PLoS One, 2012, vol. 7, no. 7. Available at: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3393741, free access (accessed: 14.03.13).
  17. Steingart K.R., Dendukuri N., Henry M., Schiller I., Nahid P., Hopewell P.C., Ramsay A., Pai M., Laal S. Performance of purified antigens for serodiagnosis of pulmonary tuberculosis: a meta-analysis. Clin. Vaccine Immunol., 2009, vol. 16, no. 2, pp. 260–276.
  18. Steingart K.R., Flores L.L., Dendukuri N., Schiller I., Laal S., Ramsay A., Hopewell P.C., Pai M. Commercial serological tests for the diagnosis of active pulmonary and extrapulmonary tuberculosis: an updated systematic review and meta-analysis. PLoS Med., 2011, vol. 8, no. 8. Available at: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3153457, free access (accessed: 14.03.13).
  19. Wallis R.S., Pai M., Menzies D., Doherty T.M., Walzl G., Perkins M.D., Zumla A. Biomarkers and diagnostics for tuberculosis: progress, needs, and translation into practice. Lancet, 2010, vol. 375, iss. 9729, pp. 1920–1937.
  20. Walzl G., Ronacher K., Hanekom W., Scriba T.J., Zumla A. Immunological biomarkers of tuberculosis. Nat. Rev. Immunol., 2011, vol. 11, no. 5, pp. 343–354.
  21. Weldingh K., Rosenkrands I., Okkels L.M., Doherty T.M., Andersen P. Assessing the serodiagnostic potential of 35 Mycobacterium tuberculosis proteins and identification of four novel serological antigens. J. Clin. Microbiol., 2005, vol. 43, no. 1, pp. 57–65.
  22. Wu X., Yang Y., Zhang J., Li B., Liang Y., Zhang C., Dong M. Comparision of antibody responses to seventeen antigens from Mycobacterium tuberculosis. Clinica Сhim. Acta., 2010, vol. 411, iss. 19–20, pp. 1520–1528.
  23. Wu X.,Yang Y., Zhang J., Li B., Liang Y., Zhang C., Dong M., Cheng H., He J. Humoral immune responses against the Mycobacterium tuberculosis 38-kilodalton, MTB48, and CFP-10/ESAT-6 antigens in tuberculosis. Clin. Vaccine Immunol., 2010, vol. 17, no. 3, pp. 372–375.
  24. Xu J.N., Chen J.P., Chen D.L.Serodiagnosis efficacy and immunogenicity of the fusion protein of Mycobacterium tuberculosis composed of the 10-kilodalton culture filtrate protein, ESAT-6, and the extracellular domain fragment of PPE68. Clin. Vaccine Immunol., 2012, vol. 19, no. 4, pp. 536–544.
  25. Zhu C., Liu J., Ling Y., Yang H., Liu Z., Zheng R., Qin L., Hu Z. Evaluation of the clinical value of ELISA based on MPT64 antibody aptamer for serological diagnosis of pulmonary tuberculosis. BMC Infect. Dis., 2012, vol. 12: 96. Available at: http://www.biomedcentral.com/1471–2334/12/96, free access (accessed: 14.03.13).

Supplementary files

Supplementary Files
Action
1. JATS XML
Download

Copyright (c) 2014 Vasilyeva E.V., Verbov V.N., Beklemishev A.B., Mamaev A.L., Tsyrul’nikov A.O., Stepanenko T.A., Ivanovskii V.B., Lapin S.V., Totolian A.A.

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.
СМИ зарегистрировано Федеральной службой по надзору в сфере связи, информационных технологий и массовых коммуникаций (Роскомнадзор).
Регистрационный номер и дата принятия решения о регистрации СМИ: серия ПИ № ФС 77 - 64788 от 02.02.2016.


This website uses cookies

You consent to our cookies if you continue to use our website.

About Cookies