transmedТрансляционная медицинаTranslational Medicine2311-44952410-5155Almazov National Medical Research Centre, Saint Petersburg, Russia10.18705/2311-4495-2014-0-3-23-28transmed-33Research ArticleМОЛЕКУЛЯРНАЯ БИОЛОГИЯ И ГЕНЕТИКАПрименение maldi-tof масс-спектрометрии в клинической микробиологииMALDI-TOF MASS SPECTROMETRY IN CLINICAL MICROBIOLOGYБаранцевичЕлена ПетровнаBarantsevichE. P.lenabara2003@inbox.ruБаранцевичНаталья ЕвгеньевнаBarantsevichN. E.noemail@neicon.ruФГБУ «Федеральный медицинский исследовательский центр им. В.А. Алмазова» Минздрава России; Первый Санкт-Петербургский государственный медицинский университет им. И.П. ПавловаРоссияFederal Almazov Medical Research Centre; I. P. Pavlov First Saint-Petersburg State Medical UniversityRussian FederationПервый Санкт-Петербургский государственный медицинский университет им. И.П. ПавловаРоссияFederal Almazov Medical Research CentreRussian Federation201423122016032328Copyright © Баранцевич Е.П., Баранцевич Н.Е., 20162016Баранцевич Е.П., Баранцевич Н.Е.Barantsevich E.P., Barantsevich N.E.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://transmed.almazovcentre.ru/jour/article/view/33

До недавнего времени микробиологическая идентификация микроорганизмов в клинических лабораториях в основном полагалась на фенотипические методы. «Золотой стандарт» идентификации микроорганизмов - анализ нуклеотидной последовательности генов «домашнего хозяйства», который был внедрен в клиническую практику с 2008 года, - не получил широкого распространения. Развитие времяпролетной масс-спектрометрии с матрично-активированной лазерной десорбцией/ионизацией (MALDI-TOF MS) революционно изменило подход к рутинной идентификации микроорганизмов в клинических микробиологических лабораториях. Этот метод отличают высокая производительность, эффективность и низкая цена. Использование стандартизированных методик MALDI-TOF MS позволяет точно идентифицировать до вида большинство клинически значимых бактерий. При применении этой технологии анализируются спектральные характеристики значительного числа белковых молекул, преимущественно рибосомальных белков, являющихся уникальным «отпечатком пальца» конкретного микроорганизма. Также надежна и рутинная идентификация дрожжей с использованием MALDI-TOF MS.

Identification of microorganisms in most clinical microbiology laboratories until recently was based predominantely on phenotypic methods. The “golden standard” of identification, which is sequencing of the “house-keeping” genes of microorganisms was introduced in practice in 2008, though still is not widespread. Development of the matrix-assisted laser desorbtion-ionization time-of-flight mass spectrometry (MALDI-TOF MS) has dramatically changed the approach to the routine identification of microorganisms. The method is characterized by high output, effectiveness and low price. Use of standard MALDI-TOF MS procedures allows correct identification to the species level in most clinically important bacteria. This technology analyses spectral characteristics of significant number of proteins, mostly ribosomal, that present the unique “fingerprint” of a specific microorganism. MALDI-TOF routine yeast identification appeared to be also highly effective.

масс-спектрометриябактериигрибыидентификацияmass spectrometrybacteriafungiidentificationReferencesAnhalt J.P., Fenselau C. Identification of bacteria using mass spectrometry // Anal. Chem. - 1975. - Vol. 47. - P. 219-225.Anhalt J.P., Fenselau C. Identification of bacteria using mass spectrometry // Anal. Chem. - 1975. - Vol. 47. - P. 219-225.Tanaka K., Fenn J.B., Wüthrich K. The Nobel Prize in Chemistry 2002 // [Nobel Media AB 2014]. 2002. - URL: http://www.nobelprize.org/nobel_prizes/chemistry/laureates/2002/popular.html (дата обращения: 16.07.2014)Tanaka K., Fenn J.B., Wüthrich K. The Nobel Prize in Chemistry 2002 // [Nobel Media AB 2014]. 2002. - URL: http://www.nobelprize.org/nobel_prizes/chemistry/laureates/2002/popular.html (дата обращения: 16.07.2014)Holland R.D., Wilkes H.G., Rafii F. et al. Rapid identification of intact whole bacteria based on spectral patterns using matrix-assisted laser desorption/ionization with time-of-flight mass spectrometry // Rapid Commun. Mass Spectrom. - 1996. - Vol. 10. - P. 1227-1232.Holland R.D., Wilkes H.G., Rafii F. et al. Rapid identification of intact whole bacteria based on spectral patterns using matrix-assisted laser desorption/ionization with time-of-flight mass spectrometry // Rapid Commun. Mass Spectrom. - 1996. - Vol. 10. - P. 1227-1232.Claydon M.A., Davey S.N., Edwards-Jones V. et al. The rapid identification of intact microorganisms using mass spectrometry // Nat. Biotechnol. - 1996. - Vol. 14. - P. 1584-1586.Claydon M.A., Davey S.N., Edwards-Jones V. et al. The rapid identification of intact microorganisms using mass spectrometry // Nat. Biotechnol. - 1996. - Vol. 14. - P. 1584-1586.Haag A.M., Taylor S.N., Johnston K.H. et al. Rapid identification and speciation of Haemophilus bacteria by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry // J. Mass Spectrom. - 1998. - Vol. 33. - P. 750-756.Haag A.M., Taylor S.N., Johnston K.H. et al. Rapid identification and speciation of Haemophilus bacteria by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry // J. Mass Spectrom. - 1998. - Vol. 33. - P. 750-756.Vargha M., Takats Z., Koponka A. et al. Optimization of MALDI-TOF MS for strain level differentiation of Arthrobacter isolates // J. Microbiol Methods. - 2006. - Vol. 66. - P. 399-409.Vargha M., Takats Z., Koponka A. et al. Optimization of MALDI-TOF MS for strain level differentiation of Arthrobacter isolates // J. Microbiol Methods. - 2006. - Vol. 66. - P. 399-409.Nagy E., Maier T., Urban E. et al. Species identification of clinical isolates of Bacteroides by matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry // Clin. Microbiol. Infect. - 2009. - Vol. 15. - P. 796-802.Nagy E., Maier T., Urban E. et al. Species identification of clinical isolates of Bacteroides by matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry // Clin. Microbiol. Infect. - 2009. - Vol. 15. - P. 796-802.Giebel R., Worden C., Rust S. M. et al. Microbial fingerprinting using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) applications and challenges // Adv. Appl. Microbiol. - 2010. - Vol. 71. - P. 149-184.Giebel R., Worden C., Rust S. M. et al. Microbial fingerprinting using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) applications and challenges // Adv. Appl. Microbiol. - 2010. - Vol. 71. - P. 149-184.Croxatto A., Prod'hom G., Greub G. Applications of MALDI-TOF mass spectrometry in clinical diagnostic microbiology // FEMS Microbiol. Rev. - 2012. - Vol. 36. - P. 380-407.Croxatto A., Prod'hom G., Greub G. Applications of MALDI-TOF mass spectrometry in clinical diagnostic microbiology // FEMS Microbiol. Rev. - 2012. - Vol. 36. - P. 380-407.Knot P.D., Fisher M.A. Mass Spectrometry in Clinical Microbiology Laboratory // Clinical Microbiology Newsletter. - 2012. - Vol. 34, № 17. - P. 135-140.Knot P.D., Fisher M.A. Mass Spectrometry in Clinical Microbiology Laboratory // Clinical Microbiology Newsletter. - 2012. - Vol. 34, № 17. - P. 135-140.Mimica M.J., Valle Martino M.D., Pasternak J. MALDI-TOF MS in the clinical microbiology laboratory // J. Bras. Patol. Med. Lab. - 2013. - Vol. 49, № 4. - P. 256-259.Mimica M.J., Valle Martino M.D., Pasternak J. MALDI-TOF MS in the clinical microbiology laboratory // J. Bras. Patol. Med. Lab. - 2013. - Vol. 49, № 4. - P. 256-259.Patel R. MALDI-TOF Mass Spectrometry: Transformative Proteomics for Clinical Microbiology // Clin. Chem. - 2013. - Vol. 59, № 2. - P. 340-342.Patel R. MALDI-TOF Mass Spectrometry: Transformative Proteomics for Clinical Microbiology // Clin. Chem. - 2013. - Vol. 59, № 2. - P. 340-342.Fenselau C., Demirev P.A. Characterization of intact microorganisms by MALDI mass spectrometry // Mass Spectrom. Rev. - 2001. - Vol. 20. - P. 157-171.Fenselau C., Demirev P.A. Characterization of intact microorganisms by MALDI mass spectrometry // Mass Spectrom. Rev. - 2001. - Vol. 20. - P. 157-171.Vaidyanathan S., Winder C.L., Wade S.C. et al. Sample preparation in matrix-assisted laser desorption/ionization mass spectrometry of whole bacterial cells and the detection of high mass (> 20 kDa) proteins // Rapid Commun. Mass Spectrom. - 2002. - Vol. 16. - P. 1276-1286.Vaidyanathan S., Winder C.L., Wade S.C. et al. Sample preparation in matrix-assisted laser desorption/ionization mass spectrometry of whole bacterial cells and the detection of high mass (> 20 kDa) proteins // Rapid Commun. Mass Spectrom. - 2002. - Vol. 16. - P. 1276-1286.Williams T.L., Andrzejewski D., Lay J.O. et al. Experimental factors affecting the quality and reproducibility of MALDI TOF mass spectra obtained from whole bacteria cells // J. Am. Soc. Mass Spectrom. - 2003. - Vol. 14. - P. 342-351.Williams T.L., Andrzejewski D., Lay J.O. et al. Experimental factors affecting the quality and reproducibility of MALDI TOF mass spectra obtained from whole bacteria cells // J. Am. Soc. Mass Spectrom. - 2003. - Vol. 14. - P. 342-351.Bizzini A., Jaton K., Romo В. et al. Matrix-assisted laser desorption ionization-time of flight mass spectrometry as an alternative to 16S rRNA gene sequencing for identification of difficult-to-identify bacterial strains // J. Clin. Microbiol. - 2011. - Vol. 49. - P. 693-696.Bizzini A., Jaton K., Romo В. et al. Matrix-assisted laser desorption ionization-time of flight mass spectrometry as an alternative to 16S rRNA gene sequencing for identification of difficult-to-identify bacterial strains // J. Clin. Microbiol. - 2011. - Vol. 49. - P. 693-696.Lavigne J.-P., Espinal P., Dunyach-Remy C. et al. Mass spectrometry: a revolution in clinical microbiology? // Clin. Chem. Lab Med. - 2013. - Vol. 51, № 2. - P. 257-270.Lavigne J.-P., Espinal P., Dunyach-Remy C. et al. Mass spectrometry: a revolution in clinical microbiology? // Clin. Chem. Lab Med. - 2013. - Vol. 51, № 2. - P. 257-270.Clark A.E., Kaleta E.J., Arora A., Wolk D.M. Matrix-Assisted Laser Desorbtion Ionization-Time of Flight Mass Specrometry : a Fundamental Shift in the Routine Practice of Clinical Microbiology // Clin. Microbiol. Rev. - 2013. - Vol. 26, № 3. - P. 547-603.Clark A.E., Kaleta E.J., Arora A., Wolk D.M. Matrix-Assisted Laser Desorbtion Ionization-Time of Flight Mass Specrometry : a Fundamental Shift in the Routine Practice of Clinical Microbiology // Clin. Microbiol. Rev. - 2013. - Vol. 26, № 3. - P. 547-603.De Carolis E., Vella A., Vaccaro L. et al. Development and Validation of an In-House Database for Matrix-Assisted Laser Desorbtion Ionization-Time of Flight Mass Spectrometry-Based Yeast Identification Using a Fast Protein Extraction Procedure // J. Clin. Microbiol. - 2014. - Vol. 52, № 5. - P. 1453-1458.De Carolis E., Vella A., Vaccaro L. et al. Development and Validation of an In-House Database for Matrix-Assisted Laser Desorbtion Ionization-Time of Flight Mass Spectrometry-Based Yeast Identification Using a Fast Protein Extraction Procedure // J. Clin. Microbiol. - 2014. - Vol. 52, № 5. - P. 1453-1458.Bizzini A., Durussel C., Bille J. et al. Performance of matrix-assisted laser desorption ionization-time of flight mass spectrometry for identification of bacterial strains routinely isolated in a clinical microbiology laboratory // J. Clin. Microbiol. - 2010. - Vol. 48. - P. 1549-1554.Bizzini A., Durussel C., Bille J. et al. Performance of matrix-assisted laser desorption ionization-time of flight mass spectrometry for identification of bacterial strains routinely isolated in a clinical microbiology laboratory // J. Clin. Microbiol. - 2010. - Vol. 48. - P. 1549-1554.Bessède E., Agla-Gre M., Delagarde Y. et al. Matrix-assisted laser-desorption/ionization biotyper: experience in the routine of a University hospital // Clin. Microbiol. Infect. - 2011. - Vol. 17. - P. 533-538.Bessède E., Agla-Gre M., Delagarde Y. et al. Matrix-assisted laser-desorption/ionization biotyper: experience in the routine of a University hospital // Clin. Microbiol. Infect. - 2011. - Vol. 17. - P. 533-538.Sogawa K., Watanabe M., Sato K. et al. Use of the MALDI BioTyper system with MALDI-TOF mass spectrometry for rapid identification of microorganisms // Anal. Bioanal. Chem. - 2011. - Vol. 400. - P. 1905-1911.Sogawa K., Watanabe M., Sato K. et al. Use of the MALDI BioTyper system with MALDI-TOF mass spectrometry for rapid identification of microorganisms // Anal. Bioanal. Chem. - 2011. - Vol. 400. - P. 1905-1911.Neville S.A., Lecordier A., Ziochos H. et al. Utility of matrix-assisted laser desorption ionization-time of flight mass spectrometry following introduction for routine laboratory bacterial identification // J. Clin. Microbiol. - 2011. - Vol. 49. - P. 2980-2984.Neville S.A., Lecordier A., Ziochos H. et al. Utility of matrix-assisted laser desorption ionization-time of flight mass spectrometry following introduction for routine laboratory bacterial identification // J. Clin. Microbiol. - 2011. - Vol. 49. - P. 2980-2984.Kostrzewa M., Sparbier K., Maier T., Schubert S. MALDI-TOF MS: an upcoming tool for rapid detection of antibiotic resistance in microorganisms // Proteomics Clin. Appl. - 2013. - Vol. 7, № 11-12. - P. 767-778.Kostrzewa M., Sparbier K., Maier T., Schubert S. MALDI-TOF MS: an upcoming tool for rapid detection of antibiotic resistance in microorganisms // Proteomics Clin. Appl. - 2013. - Vol. 7, № 11-12. - P. 767-778.Edwards-Jones V., Claydon M.A., Evanson D.J. et al. Rapid discrimination between methicillin-sensitive and methicillin resistant Staphylococcus aureus by intact cell mass spectrometry // J. Med. Microbiol. - 2000. - Vol. 49. - P. 295-300.Edwards-Jones V., Claydon M.A., Evanson D.J. et al. Rapid discrimination between methicillin-sensitive and methicillin resistant Staphylococcus aureus by intact cell mass spectrometry // J. Med. Microbiol. - 2000. - Vol. 49. - P. 295-300.Burckhardt I., Zimmermann S. Using matrix-assisted laser desorption ionization-time of flight mass spectrometry to detect carbapenem resistance within 1 to 2.5 hours // J. Clin Microbiol. - 2011. - Vol. 49. - P. 3321-3324.Burckhardt I., Zimmermann S. Using matrix-assisted laser desorption ionization-time of flight mass spectrometry to detect carbapenem resistance within 1 to 2.5 hours // J. Clin Microbiol. - 2011. - Vol. 49. - P. 3321-3324.Hooff G.P., van Kampen J.J., Meesters R.J. et al. Characterization of β-lactamase enzyme activity in bacterial lysates using MALDI-mass spectrometry // J. Proteome Res. - 2012. - Vol. 11:79. - P. 84.Hooff G.P., van Kampen J.J., Meesters R.J. et al. Characterization of β-lactamase enzyme activity in bacterial lysates using MALDI-mass spectrometry // J. Proteome Res. - 2012. - Vol. 11:79. - P. 84.Hrabak J., Walkova R., Studentova V. et al. Carbapenemase activity detection by matrix-assisted laser desorption ionization-time of flight mass spectrometry // J. Clin. Microbiol. - 2011. - Vol. 49. - P. 3222-3227.Hrabak J., Walkova R., Studentova V. et al. Carbapenemase activity detection by matrix-assisted laser desorption ionization-time of flight mass spectrometry // J. Clin. Microbiol. - 2011. - Vol. 49. - P. 3222-3227.Camara J.E., Hays F.A. Discrimination between wild-type and ampicillin-resistant Escherichia coli by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry // Anal. Bioanal. Chem. - 2007. - Vol. 389. - P. 1633-1638.Camara J.E., Hays F.A. Discrimination between wild-type and ampicillin-resistant Escherichia coli by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry // Anal. Bioanal. Chem. - 2007. - Vol. 389. - P. 1633-1638.Wybo I., De Bel A., Soetens O. et al. Differentiation of cfiA-negative and cfiA-positive Bacteroides fragilis isolates by matrix-assisted laser desorption ionization-time of flight mass spectrometry // J. Clin. Microbiol. - 2011. - Vol. 49. - P. 1961-1964.Wybo I., De Bel A., Soetens O. et al. Differentiation of cfiA-negative and cfiA-positive Bacteroides fragilis isolates by matrix-assisted laser desorption ionization-time of flight mass spectrometry // J. Clin. Microbiol. - 2011. - Vol. 49. - P. 1961-1964.Sandalakis V. Psaroulaki A., De Bock P.J. et al. Investigation of rifampicin resistance mechanisms in Brucella abortus using MS-driven comparative proteomics // J. Proteome Res. - 2012. - Vol. 11. - P. 2374-2385.Sandalakis V. Psaroulaki A., De Bock P.J. et al. Investigation of rifampicin resistance mechanisms in Brucella abortus using MS-driven comparative proteomics // J. Proteome Res. - 2012. - Vol. 11. - P. 2374-2385.Cai J. C., Hu Y.Y., Zhang R. et al. Detection of OmpK36 porin loss in Klebsiella spp. by Matrix-assisted laser desorption/ionization-time of flight mass spectrometry // J. Clin. Microbiol. - 2012. - Vol. 50. - P. 2179-2182.Cai J. C., Hu Y.Y., Zhang R. et al. Detection of OmpK36 porin loss in Klebsiella spp. by Matrix-assisted laser desorption/ionization-time of flight mass spectrometry // J. Clin. Microbiol. - 2012. - Vol. 50. - P. 2179-2182.Bittar F., Ouchenane Z., Smati F. et al. MALDI-TOF-MS for rapid detection of staphylococcal Panton-Val-entine leukocidin // Int. J. Antimicrob. Agents. - 2009. - Vol. 34. - P. 467-470.Bittar F., Ouchenane Z., Smati F. et al. MALDI-TOF-MS for rapid detection of staphylococcal Panton-Val-entine leukocidin // Int. J. Antimicrob. Agents. - 2009. - Vol. 34. - P. 467-470.Szabados F., Becker K., von Eiff C. et al. The matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS)-based protein peaks of 4448 and 5302 Da are not associated with the presence of Panton-Valentine leukocidin // Int. J. Med. Microbiol. -2011. - Vol. 301. - P. 58-63.Szabados F., Becker K., von Eiff C. et al. The matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS)-based protein peaks of 4448 and 5302 Da are not associated with the presence of Panton-Valentine leukocidin // Int. J. Med. Microbiol. -2011. - Vol. 301. - P. 58-63.Hrabak J., Chudackova E., Walkova R. Matrix-assisted laser desorption ionization time-of flight (MALDI-TOF) mass spectrometry for detection of antibiotic resistance mechanisms: from research to routine diagnosis // Clin. Microbiol. Rev. - 2013. - Vol. 26, № 1. - P. 103-114.Hrabak J., Chudackova E., Walkova R. Matrix-assisted laser desorption ionization time-of flight (MALDI-TOF) mass spectrometry for detection of antibiotic resistance mechanisms: from research to routine diagnosis // Clin. Microbiol. Rev. - 2013. - Vol. 26, № 1. - P. 103-114.Huang A.M., Newton D., Kunapuli A. et al. Impact of rapid organism identification via matrix-assisted laser desorption ionization time-of flight combined with antimicrobial stewardship team intervention in adult patients with bacteremia and candidemia // Clin. Infect. Dis. - 2013. - Vol. 57, № 9. - P. 1237-1245.Huang A.M., Newton D., Kunapuli A. et al. Impact of rapid organism identification via matrix-assisted laser desorption ionization time-of flight combined with antimicrobial stewardship team intervention in adult patients with bacteremia and candidemia // Clin. Infect. Dis. - 2013. - Vol. 57, № 9. - P. 1237-1245.Kiehntopf M., Schmerler D., Brunkhorst F.M. et al. Mass spectrometry-based protein patterns in the diagnosis of sepsis / systemic inflammatory response syndrome // Shock. - 2011. - Vol. 36. - P. 560-569.Kiehntopf M., Schmerler D., Brunkhorst F.M. et al. Mass spectrometry-based protein patterns in the diagnosis of sepsis / systemic inflammatory response syndrome // Shock. - 2011. - Vol. 36. - P. 560-569.Van Belkum A., Welker M., Erhard M. et al. Biomedical mass spectrometry in today’s and tomorrow’s clinical microbiology laboratory // J. Clin. Microbiol. - 2012. - Vol. 50. - P. 1513-1517.Van Belkum A., Welker M., Erhard M. et al. Biomedical mass spectrometry in today’s and tomorrow’s clinical microbiology laboratory // J. Clin. Microbiol. - 2012. - Vol. 50. - P. 1513-1517.Kempf M., Bakour S., Flaudrops C. et al. Rapid detection of carbapenem resistance in Acinetobacter baumannii using matrix-assisted laser desorption ionization-time of flight mass spectrometry // PLoS One. - 2012. - Vol. 7. - P. e31676.Kempf M., Bakour S., Flaudrops C. et al. Rapid detection of carbapenem resistance in Acinetobacter baumannii using matrix-assisted laser desorption ionization-time of flight mass spectrometry // PLoS One. - 2012. - Vol. 7. - P. e31676.Sparbier K., Schubert S., Weller U. et al. Matrix-assisted laser desorption ionization-time of flight mass spectrometry-based functional assay for rapid detection of resistance against ß-lactam antibiotics // J. Clin. Microbiol. - 2012. - Vol. 50. - P. 927-937.Sparbier K., Schubert S., Weller U. et al. Matrix-assisted laser desorption ionization-time of flight mass spectrometry-based functional assay for rapid detection of resistance against ß-lactam antibiotics // J. Clin. Microbiol. - 2012. - Vol. 50. - P. 927-937.Han X., Hu Q., Ding S. et al. Identification and immunological characteristics of chaperonin GroEL in Riemerella anatipestifer // Appl. Microbiol. Biotechnol. - 2012. - Vol. 93. - P. 1197-1205.Han X., Hu Q., Ding S. et al. Identification and immunological characteristics of chaperonin GroEL in Riemerella anatipestifer // Appl. Microbiol. Biotechnol. - 2012. - Vol. 93. - P. 1197-1205.Kothari A., Morgan M., Haake D.A. Emerging technologies for rapid identification of pathogens // Clin. Infect. Dis. - 2014. - Vol. 59, № 2. - P. 272-278.Kothari A., Morgan M., Haake D.A. Emerging technologies for rapid identification of pathogens // Clin. Infect. Dis. - 2014. - Vol. 59, № 2. - P. 272-278.

The authors declare that there are no conflicts of interest present.