NEW OPPORTUNITIES TO IDENTIFY AND TYPE STAPHYLOCOCCUS spp. BY USING MALDI-TOF MASS SPECTROMETRY

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Abstract

Abstract. Mass spectrometry profiles of microorganisms obtained by time-of-flight matrix-associated laser desorption/ionization (MALDI-TOF) mass spectrometry are a source of information about peptide profiles can be used for microbial identification and typing. A variety of technical and bioinformational solutions complicate developing of a united mass-spectro-profile database. Staphylococcus spp. strains are good studied objects for identification by MALDI-TOF mass spectrometry, frequently resulting in nosocomial infections, especially in immunocompromised patients. Rapid differentiation of nosocomial, multiresistant and highly virulent isolates of Staphylococcus spp. Allows to reduce the lethality in weakened and immunocompromised patients. The study was aimed at assessing comparability and reproducibility of identification and typing results for Staphylococcus spp by MALDI-TOF mass spectrometry. Comparing 292 Staphylococcus spp. isolates in clinical specimens obtained fron the multidisciplinary hospital at the NWSMU im. I.I. Mechnikov was carried out by using MALDI-TOF mass spectrometer BactoSCREEN ID (Litech, Russia) and Bruker Biotyper 3.1 (Bruker GmbH, Germany). Comparability of Staphylococcus spp. identification showed that 95.9%; 12 isolates (4.1%) by “Bruker Biotyper 3.1” and 3 isolates (1.1%) by using “BactoSCREEN ID” were incorrectly identified. Repeated identification leveled the differences between the systems used. In addition, it was shown that the method of protein extraction did not affect reliability of Staphylococcus spp. species identification by using databases (÷2, p > 0.05) compared to intraspecific typing (÷2, p < 0.0001). Using different extraction protocols showed that Staphylococcus spp. mass-spectra differed by peak intensity level within the mass range up to 4000 m/z, 5300±600 m/z and 6500±500 m/z, as well as higher than 7000 m/z. Peaks of low-molecular weight peptides were detected under full protein extraction compared to sample preparation on plate extraction. To develop a unified protocol for mass-spectrometry profile processing, a reliability of the basic statistical variables (mode, median, maximum, minimum and arithmetic mean) was evaluated. Analysis of the median mass spectrometry profiles is recommended for Staphylococcus spp. intraspecific typing by using MALDI-TOF mass spectrometry as the most reproducible and consistent approach. As a result, two systems for MALDI-TOF mass spectrometry reliably identify Staphylococcus spp., but standardization of sample preparation and bioinformation analysis is required for Staphylococcus spp. typing.

About the authors

A. S. Stepanov

North-Western State Medical University named after I.I. Mechnikov, St. Petersburg

Author for correspondence.
Email: aleksandr.stepanov@szgmu.ru

Assistant Professor, Department of Medical Microbiology

195067, Russian Federation, St. Petersburg, Piskarevsky pr., 47
Phone: +7 (921) 897-10-13 (mobile)

Russian Federation

N. V. Vasilyeva

North-Western State Medical University named after I.I. Mechnikov, St. Petersburg

Email: fake@neicon.ru
PhD, MD (Biology), Professor, Head of the Department of Medical Microbiology Russian Federation

References

  1. Гостев В.В., Калиногорская О.С., Черненькая Т.В., Науменко З.С., Ворошилова Т.М., Захарова Ю.А., Хохлова О.Е., Круглов А.Н., Сидоренко С.В. Антибиотикорезистентность метициллинорезистентных Staphylococcus aureus, циркулирующих в Российской Федерации // Антибиотики и химиотерапия. 2015. Т. 60, № 1–2. C. 3–9. [Gostev V.V., Kalinogorskaya O.S., Chernenkaya T.V., Naumenko Z.S., Voroshilova T.M., Zakharova Yu.A., Khokhlova O.E., Kruglov A.N., Sidorenko S.V. Antibiotic resistance of MRSA in Russian Federation. Antibiotiki i khimioterapiya = Antibiotics and Chemotherapy, 2015, vol. 60, no. 1–2, pp. 3–9. (In Russ.)]
  2. Козлова Н.С., Баранцевич Е.П., Баранцевич Н.Е. Антибиотикорезистентность стафилококков, выделенных из крови // Научное обозрение. 2014. № 3. C. 184–189. [Kozlova N.S., Barantsevich E.P., Barantsevich N.E. Antibiotic resistance of staphylococci isolated from blood. Nauchnoe obozrenie = Scientific Review, 2014, no. 3. pp. 184–189. (In Russ.)]
  3. Albrethsen J. Reproducibility in protein profiling by MALDI-TOF mass spectrometry. Clin. Chem., 2007, vol. 53, no. 5, pp. 852– 858. doi: 10.1373/clinchem.2006.082644
  4. Böhme K., Fernández-No I.C., Barros-Velázquez J., Gallardo J.M., Cañas B., Calo-Mata P. Comparative analysis of protein extraction methods for the identification of seafood-borne pathogenic and spoilage bacteria by MALDI-TOF mass spectrometry. Analyt. Methods, 2010, vol. 2, no. 12, pp. 1941–1947. doi: 10.1039/C0AY00457J
  5. Camoez M., Sierra J.M., Dominguez M.A., Ferrer-Navarro M., Vila J., Roca I. Automated categorization of methicillin-resistant Staphylococcus aureus clinical isolates into different clonal complexes by MALDI-TOF mass spectrometry. Clin. Microbiol. Infect., 2016, vol. 22, no. 2, pp. 161.e1–161.e7. doi: 10.1016/j.cmi.2015.10.009
  6. Chen X.-P., Li W.G., Zheng H., Du H.Y., Zhang L. Extreme diversity and multiple SCCmec elements in coagulase-negative Staphylococcus found in the Clinic and Community in Beijing, China. Ann. Clin. Microbiol. Antimicrob., 2017, vol. 16: 57. doi: 10.1186/s12941-017-0231-z
  7. Gibb S., Strimmer K. MALDIquant: a versatile R package for the analysis of mass spectrometry data. Bioinformatics (Oxford, England), 2012, vol. 28, no. 17, pp. 2270–2271. doi: 10.1093/bioinformatics/bts447
  8. Jamal W., Albert M.J., Rotimi V.O. Real-time comparative evaluation of bioMerieux VITEK MS versus Bruker Microflex MS, two matrix-assisted laser desorption-ionization time-of-flight mass spectrometry systems, for identification of clinically significant bacteria. BMC Microbiology, 2014, vol. 1, no. 14: 289. doi: 10.1186/s12866-014-0289-0
  9. Lee M., Chung HS., Moon HW., Lee SH., Lee K. Comparative evaluation of two matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) systems, Vitek MS and Microflex LT, for the identification of Grampositive cocci routinely isolated in clinical microbiology laboratories. J. Microbiol. Methods, 2015, vol. 113, pp. 13–15. doi: 10.1016/j.mimet.2015.03.020
  10. Patel R. MALDI-TOF MS for the diagnosis of infectious diseases. Clin. Chem., 2015, vol. 61, no. 1, pp. 100–111.
  11. Ryan C.G., Clayton E., Griffin W.L., Sie S.H., Cousens D.R. SNIP, a statistics-sensitive background treatment for the quantitative analysis of PIXE spectra in geoscience applications. Nucl. Instrum. Methods Phys. Res. B., 1988, vol. 34, pp. 396–402. doi: 10.1016/0168-583X(88)90063-8
  12. Schulthess B., Brodner K., Bloemberg G.V., Zbinden R., Böttger EPP., Hombach M. Identification of Gram-positive cocci by use of matrix-assisted laser desorption ionization–time of flight mass spectrometry: comparison of different preparation methods and implementation of a practical algorithm for routine diagnostics. J. Clin. Microbiol., 2013, vol. 51, no. 6, pp. 1834–1840. doi: 10.1128/JCM.02654-12
  13. Schulthess B., Bloemberg G.V., Zbinden R., Böttger E.C., Hombach M. Evaluation of the Bruker MALDI Biotyper for identification of gram-positive rods: development of a diagnostic algorithm for the clinical laboratory. J. Clin. Microbiol., 2014, vol. 52, no. 4, pp. 1089–1097. doi: 10.1128/JCM.02399-13
  14. Schuster D., Josten M., Janssen K., Bodenstein I., Albert C., Schallenberg A., Gajdiss M., Sib E., Szekat C., Kehl K., Parčina M., Hischebeth G.T.R., Bierbaum G. Detection of methicillin-resistant coagulase-negative staphylococci harboring the class A mec complex by MALDI-TOF mass-spectrometry. Int. J. Med. Microbiol., 2018, vol. 308, no. 5, pp. 522–526. doi: 10.1016/j.ijmm.2018.05.001
  15. Seng P., Fournier PE., La Scola B., Drancourt M., Raoult D. MALDI-TOF-mass spectrometry applications in clinical microbiology. Future Microbiol., 2010, vol. 5, no. 11, pp. 1733–1754. doi: 10.2217/fmb.10.127
  16. Staphylococcus aureus subtyping for MRSA detection. URL: https://www.bruker.com/products/mass-spectrometry-and-separations/maldi-biotyper-for-microbiological-research/mbt-subtyping-software-module/staphylococcus-aureus-subtyping-for-mrsa-detection.html (21.11.2017)
  17. Wang H.-Y., Lee T.Y., Tseng Y.J., Liu T.P., Huang K.Y., Chang Y.T., Chen C.H., Lu J.J. A new scheme for strain typing of methicillin-resistant Staphylococcus aureus on the basis of matrix-assisted laser desorption ionization time-of-flight mass spectrometry by using machine learning approach. PLoS One, 2018, vol. 13, no. 3, pp. e0194289. doi: 10.1371/journal.pone.0194289
  18. Westblade L.F., Garner O.B., MacDonald K., Bradford C., Pincus D.H., Mochon A.B., Jennemann R., Manji R., Bythrow M., Lewinski M.A., Burnham C.-A.D., Ginocchio C.C. Assessment of reproducibility of matrix-assisted laser desorption ionization-time of flight mass spectrometry for bacterial and yeast identification. J. Clin. Microbiol., 2015, vol. 53, no. 7, pp. 2349–2352. doi: 10.1128/JCM.00187-15
  19. Wolters M., Rohde H., Maier T., Belmar-Campos C., Franke G., Scherpe S., Aepfelbacher M., Christner M. MALDI-TOF MS fingerprinting allows for discrimination of major methicillin-resistant Staphylococcus aureus lineages. Int. J. Med. Microbiol., 2011, vol. 301, no. 1, pp. 64–68. doi: 10.1016/j.ijmm.2010.06.002

Copyright (c) 2018 Stepanov A.S., Vasilyeva N.V.

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