Download PDF
CC BY-NC-ND 4.0 · J Lab Physicians 2023; 15(03): 336-343
DOI: 10.1055/s-0042-1760401
Original Article

Role of Matrix-Assisted Laser Desorption Ionization–Time-of-Flight Mass Spectrometry for Species Identification of Acinetobacter Strains

Anusha Krishnaraj 
1   Department of Microbiology, St John's Medical College, Bengaluru, Karnataka, India
,
Pratima Gupta 
2   Department of Microbiology, All India Institute of Medical Sciences, Deoghar, Jharkhand, India
,
Mohit Bhatia
2   Department of Microbiology, All India Institute of Medical Sciences, Deoghar, Jharkhand, India
,
Balram Ji Omar 
2   Department of Microbiology, All India Institute of Medical Sciences, Deoghar, Jharkhand, India
› Author Affiliations Funding None declared.
› Further Information
   Permissions and Reprints

Abstract

Introduction Acinetobacter species has become a leading cause of nosocomial infections in recent years.

Objectives The aim of the study was to establish the usefulness of matrix-assisted laser desorption ionization–time-of-flight (MALDI-TOF) mass spectrometry (MS) for the identification of Acinetobacter species with respect to conventional biochemical methods and MicroScan WalkAway 96 Plus system and to compare the antibiotic susceptibility test results Kirby–Bauer disk diffusion method with MicroScan WalkAway 96 Plus automated identification and antimicrobial susceptibility testing system.

Materials and Methods The study sample comprised 100 clinical isolates of Acinetobacter species. They were all identified using MALDI-TOF MS and compared with other two identification systems.

Statistical Analysis Comparison of categorical variables by Fisher's exact test or Pearson's chi-square test was done. All statistical tools were two tailed, and a significant level p < 0.05 was used. All statistical tests were performed using SPSS v22.0 (Armonk IBM Corp., New York, United States). Cohen's kappa coefficients were also calculated and used as applicable.

Results MALDI-TOF MS revealed 92 A. baumannii, 2 Acinetobacter nosocomialis, 3 Acinetobacter lwoffii, and 1 each was identified as Acinetobacter junii, Acinetobacter johnsonii, and Acinetobacter tandoii. There was moderate agreement between identification by MicroScan WalkAway and MALDI-TOF, and substantial agreement between conventional biochemical tests and MALDI-TOF. We found that there was a 100% categorical agreement with respect to susceptibility of aminoglycosides (amikacin, gentamicin, tobramycin) and cephalosporins (ceftazidime, cefepime, cefotaxime) between disk diffusion method and MicroScan WalkAway 96 Plus system. Total of 16 errors were observed.

Conclusion Although MALDI-TOF MS could be useful to identify A. baumannii but not other species in the genus, it is a rapid, reliable method and can be routinely used in clinical laboratories.

Keywords

Acinetobacter species - antibiotic sensitivity - MALDI-TOF - MicroScan

Authors' Contribution

P.G., A.K., M.B., and B.J.O. conceptualized and designed the study; and definition of intellectual content, literature search, data acquisition, data analysis, statistical analysis, manuscript preparation, manuscript editing, and manuscript review were also done by them. P.G. was the guarantor.




Publication History

Article published online:
18 January 2023

© 2023. The Indian Association of Laboratory Physicians. This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/)

Thieme Medical and Scientific Publishers Pvt. Ltd.
A-12, 2nd Floor, Sector 2, Noida-201301 UP, India

 

  • References

  • 1 Gautam V, Singhal L, Ray P. Burkholderia cepacia complex: beyond Pseudomonas and Acinetobacter . Indian J Med Microbiol 2011; 29 (01) 4-12
    CrossrefPubMedGoogle Scholar
  • 2 Dijkshoorn L, Nemec A, Seifert H. An increasing threat in hospitals: multidrug-resistant Acinetobacter baumannii . Nat Rev Microbiol 2007; 5 (12) 939-951
    CrossrefPubMedGoogle Scholar
  • 3 Espinal P, Roca I, Vila J. Clinical impact and molecular basis of antimicrobial resistance in non-baumannii Acinetobacter . Future Microbiol 2011; 6 (05) 495-511
    CrossrefPubMedGoogle Scholar
  • 4 Chen S-F, Chang W-N, Lu C-H. et al. Adult Acinetobacter meningitis and its comparison with non-Acinetobacter gram-negative bacterial meningitis. Acta Neurol Taiwan 2005; 14 (03) 131-137
    PubMedGoogle Scholar
  • 5 Tsai H-Y, Cheng A, Liu C-Y. et al. Bacteremia caused by Acinetobacter junii at a medical center in Taiwan, 2000-2010. Eur J Clin Microbiol Infect Dis 2012; 31 (10) 2737-2743
    CrossrefPubMedGoogle Scholar
  • 6 Henao-Martínez AF, González-Fontal GR, Johnson S. A case of community-acquired Acinetobacter junii-johnsonii cellulitis. Biomedica 2012; 32 (02) 179-181
    PubMedGoogle Scholar
  • 7 Castellanos Martínez E, Telenti Asensio M, Rodríguez Blanco VM, Rodríguez Suárez ML, Morena Torrico A, Cortina Llosa A. Infective endocarditis of an interventricular patch caused by Acinetobacter haemolyticus . Infection 1995; 23 (04) 243-245
    CrossrefPubMedGoogle Scholar
  • 8 Vijayakumar S, Biswas I, Veeraraghavan B. Accurate identification of clinically important Acinetobacter spp.: an update. Future Sci OA 2019; 5 (06) FSO395
    CrossrefPubMedGoogle Scholar
  • 9 Kishii K, Kikuchi K, Matsuda N. et al. Evaluation of matrix-assisted laser desorption ionization-time of flight mass spectrometry for species identification of Acinetobacter strains isolated from blood cultures. Clin Microbiol Infect 2014; 20 (05) 424-430
    CrossrefPubMedGoogle Scholar
  • 10 La Scola B, Gundi VAKB, Khamis A, Raoult D. Sequencing of the rpoB gene and flanking spacers for molecular identification of Acinetobacter species. J Clin Microbiol 2006; 44 (03) 827-832
    CrossrefPubMedGoogle Scholar
  • 11 Chang HC, Wei YF, Dijkshoorn L, Vaneechoutte M, Tang CT, Chang TC. Species-level identification of isolates of the Acinetobacter calcoaceticus-Acinetobacter baumannii complex by sequence analysis of the 16S-23S rRNA gene spacer region. J Clin Microbiol 2005; 43 (04) 1632-1639
    CrossrefPubMedGoogle Scholar
  • 12 Bizzini A, Jaton K, Romo D, Bille J, Prod'hom G, Greub G. 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; 49 (02) 693-696
    CrossrefPubMedGoogle Scholar
  • 13 Wiwing V, Lugito NP. Antimicrobial susceptibility of multidrug-resistant Acinetobacter baumanii in a teaching hospital: A two-year observation. Open Journal of Medical Microbiology 2015; 05 (02) 85-89
    CrossrefGoogle Scholar
  • 14 Dimple R, Nupur S, Mahawal BS, Ankit K, Ajay P. Speciation and antibiotic resistance pattern of Acinetobacter species in a tertiary care hospital in Uttarakhand. International Journal of Medical Research & Health Sciences 2016; 5 (04) 89-96
    Google Scholar
  • 15 Tripathi PC, Gajbhiye SR, Agrawal GN. Clinical and antimicrobial profile of Acinetobacter spp.: an emerging nosocomial superbug. Adv Biomed Res 2014; 3: 13
    CrossrefPubMedGoogle Scholar
  • 16 Rajmane VS, Rajmane ST, Mohite ST. Study of Incidence, Risk Factors and Antibiotic Sensitivity Pattern of Acinetobacter baumannii in a Tertiary Care Hospital. Journal of Krishna Institute of Medical Sciences (JKIMSU) 2015; 4 (01) 5
    Google Scholar
  • 17 Kaur TA, Putatunda CH, Oberoi AR, Vyas AS, Kumar GA. Prevalence and drug resistance in Acinetobacter sp. isolated from intensive care units patients in Punjab, India. Asian J Pharm Clin Res 2018; 11 (14) 88-93
    CrossrefGoogle Scholar
  • 18 Babay HA, Manneh K, Somily AM. Accuracy of detecting resistance to carbapenems among gram negative rods: comparison of three methods. Journal of Taibah University Medical Sciences 2009; Jan 1; 4 (01) 53-61
    CrossrefGoogle Scholar
  • 19 Lee SY, Shin JH, Kim SH, Shin MG, Suh SP, Ryang DW. Evaluation of matrix-assisted laser desorption ionization-time of flight mass spectrometry-based VITEK MS system for the identification of Acinetobacter species from blood cultures: comparison with VITEK 2 and MicroScan systems. Ann Lab Med 2015; 35 (01) 62-68
    CrossrefPubMedGoogle Scholar
  • 20 Hsueh P-R, Kuo L-C, Chang T-C. et al. Evaluation of the Bruker Biotyper matrix-assisted laser desorption ionization-time of flight mass spectrometry system for identification of blood isolates of Acinetobacter species. J Clin Microbiol 2014; 52 (08) 3095-3100
    CrossrefPubMedGoogle Scholar
  • 21 Espinal P, Seifert H, Dijkshoorn L, Vila J, Roca I. Rapid and accurate identification of genomic species from the Acinetobacter baumannii (Ab) group by MALDI-TOF MS. Clin Microbiol Infect 2012; 18 (11) 1097-1103
    CrossrefPubMedGoogle Scholar
  • 22 Šedo O, Nemec A, Křížová L, Kačalová M, Zdráhal Z. Improvement of MALDI-TOF MS profiling for the differentiation of species within the Acinetobacter calcoaceticus-Acinetobacter baumannii complex. Syst Appl Microbiol 2013; 36 (08) 572-578
    CrossrefPubMedGoogle Scholar
  • 23 Hernández-Durán M, López-Jácome LE, Colín-Castro CA, Cerón-González G, Ortega-Peña S, Vanegas-Rodríguez ES, Mondragón-Eguiluz JA, Franco-Cendejas R. Comparison of the microscan walkaway and Vitek 2 compact systems for the identification and susceptibility of clinical gram-positive and gram-negative bacteria. Investigación en discapacidad 2017; Oct 16; 6 (03) 105-114
    Google Scholar
  • 24 Linde H-J, Hahn J, Holler E, Reischl U, Lehn N. Septicemia due to Acinetobacter junii . J Clin Microbiol 2002; 40 (07) 2696-2697
    CrossrefPubMedGoogle Scholar
  • 25 Karah N, Haldorsen B, Hegstad K, Simonsen GS, Sundsfjord A, Samuelsen Ø. Norwegian Study Group of Acinetobacter. Species identification and molecular characterization of Acinetobacter spp. blood culture isolates from Norway. J Antimicrob Chemother 2011; 66 (04) 738-744
    CrossrefPubMedGoogle Scholar
  • 26 Kulah C, Aktas E, Comert F, Ozlu N, Akyar I, Ankarali H. Detecting imipenem resistance in Acinetobacter baumannii by automated systems (BD Phoenix, MicroScan WalkAway, VITEK 2); high error rates with MicroScan WalkAway. BMC Infect Dis 2009; 9 (01) 30
    CrossrefPubMedGoogle Scholar
  • 27 Aybey AD, Aksit F, Oz Y, Kiremitci A, Durmaz G. Evaluation of an automated system for the detection of carbapenem resistant Acinetobacter baumannii and assessment of metallo-β-lactamase production using two different phenotyping methods. J Microbiol Methods 2011; 86 (01) 121-123
    CrossrefPubMedGoogle Scholar
  • 28 Magiorakos A-P, Srinivasan A, Carey RB. et al. Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clin Microbiol Infect 2012; 18 (03) 268-281
    CrossrefPubMedGoogle Scholar
  • 29 Oberoi A, Aggarwal A, Lal M. A Decade of an Underestimated Nosocomal Pathogen-Acinetobacter In a Tertiary Care Hospital in Punjab. JK science 2009 Jan 1;11(01):
    Google Scholar
  • 30 Falagas ME, Rafailidis PI, Matthaiou DK. Resistance to polymyxins: mechanisms, frequency and treatment options. Drug Resist Updat 2010; 13 (4-5): 132-138
    CrossrefPubMedGoogle Scholar
  • 31 Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing. CLSI supplement M100. 2017. Jan 18
    Google Scholar
  • 32 European Society of Clinical Microbiology. EUCAST warnings Concerning Antimicrobial Susceptibility Testing Products or Procedures [Internet]. EUCAST [accessed May 27, 2020]; at: https://www.eucast.org/ast_of_bacteria/warnings/
    Google Scholar
  • 33 Jayol A, Nordmann P, André C, Poirel L, Dubois V. Evaluation of three broth microdilution systems to determine colistin susceptibility of Gram-negative bacilli. J Antimicrob Chemother 2018; 73 (05) 1272-1278
    CrossrefPubMedGoogle Scholar
  • 34 Singh RI, Bhatia M, Anusha KR, Singh V, Omar BJ, Gupta P. Comparative evaluation of MicroScan WalkAway 96 Plus ID/AST system and Mikrolatest broth microdilution kit in assessing In vitro colistin susceptibility of carbapenem-resistant clinical gram-negative bacterial isolates: experience from a tertiary care teaching hospital in Rishikesh, Uttarakhand. Indian J Med Microbiol 2019; 37 (04) 502-508
    CrossrefPubMedGoogle Scholar