APPLICATION OF TIME-OF-FLIGHT MASS-SPECTROMETRY FOR DETECTION OF CAUSATIVE AGENT OF BRUCELLOSIS IN BLOOD SAMPLES IN EXPERIMENT

Cover Page


Cite item

Full Text

Abstract

Aim.Study the possibility to apply time-of-flight mass-spectrometry for detection of causative agent of brucellosis in blood. Materials and methods. Brucella strains: 5 Brucella melitensis and 21 Brucella abortus. Protein profiling in linear mode on MALD1-TOF mass-spectrometer Microflex «Bruker Daltonics». Results. Technique for disinfection and preparation of blood samples was modified and optimized for MALDI-TOF MS analysis. 120 representative protein profiles of sera extract were obtained that contain brucellosis causative agent. A resulting peak-list (super-spectrum) of the studied protein fraction of blood extract of a conditionally healthy human within the studied group was formed and analyzed. Conclusion. A scheme of brucella detection in blood samples by MALDI-TOF MS is proposed, based on detection of a complex of 15 genus-specific fragments. Signals on mass-spectra of extracts of leukocyte fraction of blood, artificially contaminated with brucellosis causative agents are characterized.

About the authors

D. V. Ulshina

Stavropol Research Institute for Plague Control

Author for correspondence.
Email: noemail@neicon.ru
Russian Federation

D. A. Kovalev

Stavropol Research Institute for Plague Control

Email: noemail@neicon.ru
Russian Federation

D. G. Ponomarenko

Stavropol Research Institute for Plague Control

Email: noemail@neicon.ru
Russian Federation

D. V. Rusanova

Stavropol Research Institute for Plague Control

Email: noemail@neicon.ru
Russian Federation

N. M. Shvetsova

Stavropol Research Institute for Plague Control

Email: noemail@neicon.ru
Russian Federation

T. V. Taran

Stavropol Research Institute for Plague Control

Email: noemail@neicon.ru
Russian Federation

I. V. Kuznetsova

Stavropol Research Institute for Plague Control

Email: noemail@neicon.ru
Russian Federation

A. M. Zhirov

Stavropol Research Institute for Plague Control

Email: noemail@neicon.ru
Russian Federation

A. A. Khachaturova

Stavropol Research Institute for Plague Control

Email: noemail@neicon.ru
Russian Federation

I. Yu. Borzdova

Stavropol Research Institute for Plague Control

Email: noemail@neicon.ru
Russian Federation

A. N. Kulichenko

Stavropol Research Institute for Plague Control

Email: noemail@neicon.ru
Russian Federation

References

  1. Ломинадзе Г.Г., Семенова Е.А., Мотузова О.В., Калакуцкая А.Н., Лазарева А.В. Использование метода MALDI-TOF масс-спектрометрии для ускорения идентификации микроорганизмов в гемокультурах пациентов с подозрением на сепсис. Лаборатория ЛПУ. 2014,4: 17-20.
  2. Лямкин Г.И., Пономаренко Д.Г., Худолеев А.А., Русанова Д.В., Вилинская С.В., Куличенко А.Н. Обзор эпидемиологической ситуации по бруцеллезу в Российской Федерации в 2015 г. и прогноз на 2016 г. Проблемы особо опасных инфекций. 2016, 2: 11-13.
  3. Ульшина Д.В., Ковалев Д.А., Бобрышева О.В., Лямкин Г.И., Худолеев А.А., Сирица Ю.В., Куличенко А.Н. Разработка алгоритма идентификации культур возбудителя бруцеллеза методом MALDI-TOF масс-спектрометрии. Проблемы особо опасных инфекций. 2015, 4: 96-99.
  4. Baranov А.А., Mayanskii A.N., Mayanskii N.A. A new epoch in medical microbiology. Her. Russ. Acad. Sci. 2015, 85 (6): 515-522.
  5. Ferreira L., Castano S.V., Sanchez-Juanes F. et al. Identification of Brucella by MALDI-TOF mass spectrometry. Fast and reliable identification from agar plates and blood cultures. PLoS One. 2010,5 (12): 14235.
  6. GrunowR., Jacob D., Klee S. et al. Brucellosis in a refugee who migrated from Syria to Germany and lessons learnt. Euro Surveill. 2016, 21 (31): 1-4.
  7. Karger A., Melzer F., Tirnke M. et al. Interlaboratory comparison ofintact-cell matrix-assisted laser desorption ionization-time of flight mass spectrometry results for identification and differentiation of Brucella spp. J. Clin. Microbiol. 2013, 51 (9): 3123-3126.
  8. Lasch P., Wahab T., Weil S. et al. Identification of highly pathogenic microorganisms by matrixassisted laser desorption ionization-time of flight mass spectrometry: Results of an interlaboratory ring trial. J. Clin. Microbiol. 2015, 53 (8): 2632-2640.
  9. Lista F., Reubsaet F., De Santis R. et al. Reliable identification at the species level of Brucella isolates with MALDI-TOF-MS. BMC Microbiology. 2011, 11: 267.
  10. Nyvang H.G., Kvistholm J.A., Bocher S. et al. Mass spectrometry: pneumococcal meningitis verified and Brucella species identified in less than half an hour. Scand. J. Infect. Dis. 2010, 42 (9): 716-718. v
  11. Sedo O., Sedlacek I., Zdrahal Z. Sample preparation methods for MALDI-MS profiling of bacteria. Mass Spectrometry Reviews. 2011, 30 (3): 417-434.
  12. Vitr M.A., Mambres D.H., Deghelt M. et al. Brucella melitensis invades murine erythrocytes during infection. Infect. Immunity. 2014, 82 (9): 3927-3938.
  13. Yonetani S., Olmishi H., Ohkusu K. et al. Direct identification of microorganisms from positive blood cultures by MALDI-TOF MS using an in-house saponin method. Intern. J. Infect. Dis. 2016, 52: 37-42.
  14. Zamanian M., Hashemi Tabar G.R., Rad M. et al. Evaluation of different primers for detection of Brucella in human and animal serum samples by using PCR method. Archives Iranian Medicine. 2015, 18 (1): 44-50.

Copyright (c) 2017 Ulshina D.V., Kovalev D.A., Ponomarenko D.G., Rusanova D.V., Shvetsova N.M., Taran T.V., Kuznetsova I.V., Zhirov A.M., Khachaturova A.A., Borzdova I.Y., Kulichenko A.N.

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: ПИ № ФС77-75442 от 01.04.2019 г.


This website uses cookies

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

About Cookies