Skip to main content

Advertisement

SpringerLink
Log in

Rapid colorimetric polymyxin B microelution directly from positive blood bottles: because patients with serious infections should not have to wait for results of culture-based methodologies

  • Research
  • Published:
European Journal of Clinical Microbiology & Infectious Diseases Aims and scope Submit manuscript

Abstract

Purpose

To evaluate the performance of the rapid colorimetric polymyxin B microelution (RCPEm) in determining polymyxin B resistance directly from Enterobacterales-positive blood cultures.

Methods

A set volume of positive blood culture bottles (diluted 1:10) was inoculated into a glucose-broth-phenol red solution (NP solution), where a polymyxin B disk was previously eluted (final concentration of 3 µg/mL). Test was read each 1 h for up to 4 h. Color change from red/orange to yellow indicated resistant isolates. Results were compared to the reference method, broth microdilution (BMD), performed from colonies grown on solid media from the same blood culture bottle.

Results

One hundred fifty-two Enterobacterales-positive blood cultures were evaluated, 22.4% (34/152) of them resistant to polymyxin B (including 6.6% with borderline MICs). When performing directly from positive blood cultures (RCPEm-BC), specificity and sensitivity were 99.1% and 94.1%, respectively. Of note, 79.4% (27/34) of truly resistant isolates required 3 h of incubation, compared to the 18 ± 2 h incubation that microtiter plates of BMD demand before reading can be performed.

Conclusions

RCPEm directly from blood cultures has great potential to be part of the routine of clinical microbiology laboratories to establish polymyxin B susceptibility, impacting outcome of patients with bloodstream infections caused by carbapenem-resistant Enterobacterales.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

Data availability

The datasets generated and analyzed during the current study are available from the corresponding author on reasonable request.

References

  1. Xu L, Sun X, Ma X (2017) Systematic review and meta-analysis of mortality of patients infected with carbapenem-resistant Klebsiella pneumoniae. Ann Clin Microbiol Antimicrob 16(1):18. https://doi.org/10.1186/s12941-017-0191-3

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Tsuji BT, Pogue JM, Zavascki AP et al (2019) International Consensus guidelines for the optimal use of the polymyxins: endorsed by the American College of Clinical Pharmacy (ACCP), European Society of Clinical Microbiology and Infectious diseases (ESCMID), Infectious Diseases Society of America (IDSA), International Society for anti-infective pharmacology (ISAP), society of critical Care Medicine (SCCM), and Society of Infectious diseases pharmacists (SIDP). Pharmacother J Hum Pharmacol Drug Ther 39(1):10–39. https://doi.org/10.1002/phar.2209

    Article  Google Scholar 

  3. Zeng M, Xia J, Zong Z, Shi Y et al (2023) Guidelines for the diagnosis, treatment, prevention and control of infections caused by carbapenem-resistant gram-negative bacilli. J Microbiol Immunol Infect 56(4):653–671. https://doi.org/10.1016/j.jmii.2023.01.017

    Article  CAS  PubMed  Google Scholar 

  4. Paul M, Carrara E, Retamar P et al (2022) European Society of Clinical Microbiology and Infectious diseases (ESCMID) guidelines for the treatment of infections caused by multidrug-resistant Gram-negative bacilli (endorsed by European society of intensive care medicine). Clin Microbiol Infect 28(4):521–547. https://doi.org/10.1016/j.cmi.2021.11.025

    Article  CAS  PubMed  Google Scholar 

  5. Soman R, Bakthavatchalam YD, Nadarajan A et al (2021) Is it time to move away from polymyxins? Evidence and alternatives. Eur J Clin Microbiol Infect Dis 40(3):461–475. https://doi.org/10.1007/s10096-020-04053-w

    Article  CAS  PubMed  Google Scholar 

  6. Li Z, Cao Y, Yi L, Liu JH, Yang Q (2019) Emergent Polymyxin Resistance: end of an era? Open Forum Infect Dis 6(10):ofz368. https://doi.org/10.1093/ofid/ofz368

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Silva KED, Rossato L, Leite AF, Simionatto S (2022) Overview of polymyxin resistance in Enterobacteriaceae. Rev Soc Bras Med Trop 55:e0349–2021. https://doi.org/10.1590/0037-8682-0349-2021

    Article  PubMed  PubMed Central  Google Scholar 

  8. Kadri SS, Lai YL, Warner S et al (2021) Inappropriate empirical antibiotic therapy for bloodstream infections based on discordant in-vitro susceptibilities: a retrospective cohort analysis of prevalence, predictors, and mortality risk in US hospitals. Lancet Infect Dis 21(2):241–251. https://doi.org/10.1016/S1473-3099(20)30477-1

    Article  CAS  PubMed  Google Scholar 

  9. Nordmann P, Jayol A, Poirel L (2016) Rapid Detection of Polymyxin Resistance in Enterobacteriaceae. Emerg Infect Dis 22(6):1038–1043. https://doi.org/10.3201/eid2206.151840

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Barth PO, Volpato FCZ, Moreira NK, Wink PL, De Souza ÂC, Barth AL (2022) Evaluation of a rapid susceptibility test of polymyxin B by MALDI-TOF. Front Microbiol 13:1075650. https://doi.org/10.3389/fmicb.2022.1075650

    Article  PubMed  PubMed Central  Google Scholar 

  11. Collar GdaS, Raro OHF, da Silva RMC et al (2021) Polymyxin NP tests (from colonies and directly from blood cultures): accurate and rapid methodologies to detect polymyxin B susceptibility among Enterobacterales. Diagn Microbiol Infect Dis 99(4):115264. https://doi.org/10.1016/j.diagmicrobio.2020.115264

    Article  CAS  PubMed  Google Scholar 

  12. Fonseca E, Silva D, Silva-Dias A, Gomes R, Martins-Oliveira I, Ramos MH, Rodrigues AG, Cantón R, Pina-Vaz C (2019) Evaluation of rapid colistin susceptibility directly from positive blood cultures using a flow cytometry assay. Int J Antimicrob Agents 54(6):820–823. https://doi.org/10.1016/j.ijantimicag.2019.08.016

    Article  CAS  Google Scholar 

  13. Jayol A, Dubois V, Poirel L, Nordmann P (2016) Rapid Detection of Polymyxin-Resistant Enterobacteriaceae from Blood Cultures. Bourbeau P, editor. J Clin Microbiol. 54(9):2273–7. https://doi.org/10.1128/JCM.00918-16

  14. Collar GDS, Moreira NK, Wink PL, Barth AL, Raro OHF, Dias C, De Lima Machado A, Mott MP, Caierão J (2023) Detection of polymyxins resistance among Enterobacterales: evaluation of available methods and proposal of a new rapid and feasible methodology. Ann Clin Microbiol Antimicrob 22(1):71. https://doi.org/10.1186/s12941-023-00618-7

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. ISO (2019) ISO 20776-1:2019 susceptibility testing of infectious agents and evaluation of performance of antimicrobial susceptibility test devices – part 1: broth micro-dilution reference method for testing the in vitro activity of antimicrobial agents against rapid. Geneva, Switzerland. https://www.iso.org/standard/70464.html

    Google Scholar 

  16. Barth PO, Roesch EW, Lutz L, De Souza ÂC, Goldani LZ, Pereira DC (2023) Rapid bacterial identification by MALDI-TOF MS directly from blood cultures and rapid susceptibility testing: a simple approach to reduce the turnaround time of blood cultures. Braz J Infect Dis 27(1):102721. https://doi.org/10.1016/j.bjid.2022.102721

    Article  CAS  PubMed  Google Scholar 

  17. European Committee on Antimicrobial Susceptibility Testing (EUCAST) (2024) Methodology - EUCAST rapid antimicrobial susceptibility testing (RAST) directly from blood cultures bottles. https://www.eucast.org/. Accessed 25 March 2024

  18. European Committee on Antimicrobial Susceptibility Testing (EUCAST) (2024) Antimicrobial susceptibility testing EUCAST disk diffusion method. https://www.eucast.org/. Accessed 18 March 2024

  19. European Committee on Antimicrobial Susceptibility Testing (EUCAST) (2024) Breakpoint tables for interpretation of MICs and zone diameters. https://www.eucast.org/. Accessed 30 January 2024

  20. Brazilian Committee on Antimicrobial Susceptibility Testing (BrCAST) (2023) Breakpoint tables for interpretation of MICs and zone diameters. https://www.brcast.org/. Accessed 30 January 2024

  21. Tan X, Kim HS, Baugh K, Huang Y, Kadiyala N, Wences M, Singh N, Wenzler E, Bulman ZP (2021) Therapeutic options for Metallo-β-Lactamase-Producing Enterobacterales. Infect Drug Resist Volume 14:125–142. https://doi.org/10.2147/IDR.S246174

    Article  CAS  Google Scholar 

  22. European Committee on Antimicrobial Susceptibility Testing (EUCAST) (2016) Recommendations for MIC determination of colistin (polymyxin E) As recommended by the joint CLSI-EUCAST Polymyxin Breakpoints Working Group, https://www.eucast.org. Accessed 15 February 2024

  23. Wink PL, Martins AS, Inamine E, Dalmolin TV, Barth AL (2019) Rapid detection of the main carbapenemases in Brazil directly from spiked blood culture using the RESIST-3 O.K.N. immunoassay. Braz J Microbiol 50(3):657–662. https://doi.org/10.1007/s42770-019-00109-y

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Malli E, Papagiannitsis CC, Xitsas S, Tsilipounidaki K, Petinaki E (2019) Implementation of the Rapid Polymyxin™ NP test directly to positive blood cultures bottles. Diagn Microbiol Infect Dis 95(4):114889. https://doi.org/10.1016/j.diagmicrobio.2019.114889

    Article  CAS  PubMed  Google Scholar 

  25. Luo Q, Xu L, Wang Y, Fu H, Xiao T, Yu W, Zhou W, Zhang K, Shen J, Ji J, Ying C, Xiao Y (2024) Clinical relevance, mechanisms, and evolution of polymyxin B heteroresistance carbapenem-resistant Klebsiella pneumoniae: a genomic, retrospective cohort study. Clin Microbiol Infect S1198743X:24000363. https://doi.org/10.1016/j.cmi.2024.01.014

    Article  CAS  Google Scholar 

  26. Food and Drug Administration (FDA) (2018) Antimicrobial Susceptibility Test (AST) Systems - Class II Special Controls Guidance for Industry and FDA, https://www.fda.gov. Accessed 20 February 2024

  27. Zayet S, Lang S, Garnier P, Pierron A, Plantin J, Toko L, Royer PY, Villemain M, Klopfenstein T, Gendrin V (2021) Leclercia adecarboxylata as Emerging Pathogen in Human infections: clinical features and Antimicrobial susceptibility testing. Pathogens 10(11):1399. https://doi.org/10.3390/pathogens10111399

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Funding

This study was funded by Instituto Nacional de Pesquisa em Resistência Antimicrobiana – INPRA (INCT/CNPq 465718/2014-0) and Fundo de Incentivo à Pesquisa e Eventos from Hospital de Clínicas de Porto Alegre – FIPE/ HCPA (project number 2023-0061).

Author information

Authors and Affiliations

  1. Laboratório de Pesquisa em Bacteriologia Clínica (LaBaC), Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil

    Gabriela da Silva Collar, Julia Becker, Natália Kehl Moreira & Juliana Caierão

  2. Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil

    Gabriela da Silva Collar, Natália Kehl Moreira & Juliana Caierão

  3. Laboratório de Microbiologia Clínica, Hospital de Clínicas de Porto Alegre, Porto Alegre, Brazil

    Luana Silva Dornelles & Mariana Preussler Mott

  4. Laboratório de Pesquisa em Resistência Bacteriana (LABRESIS), Hospital de Clínicas de Porto Alegre, Porto Alegre, Brazil

    Afonso Luís Barth

Authors
  1. Gabriela da Silva Collar
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Julia Becker
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Natália Kehl Moreira
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Luana Silva Dornelles
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Mariana Preussler Mott
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Afonso Luís Barth
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Juliana Caierão
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Writing—original draft: G.S.C.; writing—review and editing: G.S.C., M.P.M., A.L.B. and J.C.; conceptualization: G.S.C.; investigation: G.S.C.; methodology: G.S.C., J.B., N.K.M., L.S.D., M.P.M.; formal analysis: G.S.C.; project administration: J.C.; funding acquisition: A.L.B. and J.C.

Corresponding author

Correspondence to Gabriela da Silva Collar.

Ethics declarations

Ethics approval

Approved by Hospital de Clínicas de Porto Alegre (HCPA) from Universidade Federal do Rio Grande do Sul (UFRGS) ethical committee, under CAAE number 67871323.2.0000.5327.

Competing interests

The authors declare no competing interests.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Collar, G.d.S., Becker, J., Moreira, N.K. et al. Rapid colorimetric polymyxin B microelution directly from positive blood bottles: because patients with serious infections should not have to wait for results of culture-based methodologies. Eur J Clin Microbiol Infect Dis (2024). https://doi.org/10.1007/s10096-024-04846-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s10096-024-04846-3

Keywords

Search

Navigation