Direct microorganism species identification and antimicrobial susceptibility tests from positive blood culture bottles using rapid Sepsityper Kit

Published:January 10, 2022DOI:https://doi.org/10.1016/j.jiac.2021.12.030

      Abstract

      Introduction

      We evaluated the performance of Rapid Sepsityper Kit in species identification (ID) and antimicrobial susceptibility testing (AST).

      Methods

      Positive blood culture bottles (n = 227) containing single microorganisms were enrolled. We compared the direct method using Rapid Sepsityper Kit for ID and AST with the conventional method. The analyses of ID and AST were performed using MALDI Biotyper and BD Phoenix platform, respectively.

      Results

      The direct ID method correctly identified 89.4% (203/227) of samples, and Gram-negative bacilli (95.2%) had a higher ID rate than Gram-positive cocci (84.4%). Five cases were misidentified, and non-acceptable identification was high among Streptococcus species.
      Direct AST results were obtained from 168 isolates. Non-acceptable ID occurred among 24 isolates; 4 Streptococcus species, and 31 isolates, which did not grow in the direct AST method, were excluded. A total of 1714 antibiotic susceptibility tests (625 from 69 Gram-positive cocci and 1089 from 99 Gram-negative bacilli) were performed. The direct AST methods showed 98.3% (1685/1714) of categorical agreement (CA), 0.7% (12/1714) of very major errors, 0.2% (4/1714) of major errors, and 0.8% (13/1714) of minor errors. Complete CA was obtained for methicillin-resistant Staphylococcus aureus and extended-spectrum beta-lactamase-producing Escherichia coli.

      Conclusions

      The direct ID method using Rapid Sepsityper Kit and the direct AST method in combination with the BD Phoenix platform, which was associated with a reduction of turnaround time, may be a reliable approach for blood culture bottles. However, additional validation and further improvements, especially for Gram-positive cocci, would have an impact on microbiological diagnoses.

      Keywords

      Abbreviations:

      ID (species identification), AST (antimicrobial susceptibility testing), BSI (bloodstream infection)
      To read this article in full you will need to make a payment

      Subscribe:

      Subscribe to Journal of Infection and Chemotherapy
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Kumar A.
        • Roberts D.
        • Wood K.E.
        • Light B.
        • Parrillo J.E.
        • Sharma S.
        • et al.
        Duration of hypotension before initiation of effective antimicrobial therapy is the critical determinant of survival in human septic shock*.
        Crit Care Med. 2006; 34: 1589-1596https://doi.org/10.1097/01.Ccm.0000217961.75225.E9
        • Opota O.
        • Croxatto A.
        • Prod'hom G.
        • Greub G.
        Blood culture-based diagnosis of bacteraemia: state of the art.
        Clin Microbiol Infect. 2015; 21: 313-322https://doi.org/10.1016/j.cmi.2015.01.003
        • Rhodes A.
        • Evans L.E.
        • Alhazzani W.
        • Levy M.M.
        • Antonelli M.
        • Ferrer R.
        • et al.
        Surviving sepsis campaign: international guidelines for management of sepsis and septic shock.
        Crit Care Med. 2016; 2017: 486-552https://doi.org/10.1097/ccm.0000000000002255
        • Angeletti S.
        Matrix assisted laser desorption time of flight mass spectrometry (MALDI-TOF MS) in clinical microbiology.
        J Microbiol Methods. 2017; 138: 20-29https://doi.org/10.1016/j.mimet.2016.09.003
        • Nomura F.
        • Tsuchida S.
        • Murata S.
        • Satoh M.
        • Matsushita K.
        Mass spectrometry-based microbiological testing for blood stream infection.
        Clin Proteonomics. 2020; 17: 14https://doi.org/10.1186/s12014-020-09278-7
        • Patel T.S.
        • Kaakeh R.
        • Nagel J.L.
        • Newton D.W.
        • Stevenson J.G.
        • Bourbeau P.
        Cost analysis of implementing matrix-assisted laser desorption ionization–time of flight mass spectrometry Plus real-time antimicrobial stewardship intervention for bloodstream infections.
        J Clin Microbiol. 2017; 55: 60-67https://doi.org/10.1128/JCM.01452-16
        • Riederer K.
        • Cruz K.
        • Shemes S.
        • Szpunar S.
        • Fishbain J.T.
        MALDI-TOF identification of Gram-negative bacteria directly from blood culture bottles containing charcoal: Sepsityper® kits versus centrifugation–filtration method.
        Diagn Microbiol Infect Dis. 2015; 82: 105-108https://doi.org/10.1016/j.diagmicrobio.2015.03.003
        • Ponderand L.
        • Pavese P.
        • Maubon D.
        • Giraudon E.
        • Girard T.
        • Landelle C.
        • et al.
        Evaluation of Rapid Sepsityper® protocol and specific MBT-Sepsityper module (Bruker Daltonics) for the rapid diagnosis of bacteremia and fungemia by MALDI-TOF-MS.
        Ann Clin Microbiol Antimicrob. 2020; 60: 19https://doi.org/10.1186/s12941-020-00403-w
        • Wu S.
        • Xu J.
        • Qiu C.
        • Xu L.
        • Chen Q.
        • Wang X.
        Direct antimicrobial susceptibility tests of bacteria and yeasts from positive blood cultures by using serum separator gel tubes and MALDI–TOF MS.
        J Microbiol Methods. 2019; 157: 16-20https://doi.org/10.1016/j.mimet.2018.12.011
        • Yuan Y.
        • Wang J.
        • Zhang J.
        • Ma B.
        • Gao S.
        • Li Y.
        • et al.
        Evaluation of an optimized method to directly identify bacteria from positive blood cultures using MALDI-TOF mass spectrometry.
        J Clin Lab Anal. 2020; 34e23119https://doi.org/10.1002/jcla.23119
        • Maelegheer K.
        • Nulens E.
        Same-day identification and antibiotic susceptibility testing on positive blood cultures: a simple and inexpensive procedure.
        Eur J Clin Microbiol Infect Dis. 2017; 36: 681-687https://doi.org/10.1007/s10096-016-2849-8
        • Lee J.E.
        • Jo S.J.
        • Park K.G.
        • Suk H.S.
        • Ha S.I.
        • Shin J.S.
        • et al.
        Evaluation of modified saponin preparation method for the direct identification and antimicrobial susceptibility testing from positive blood culture.
        J Microbiol Methods. 2018; 154: 118-123https://doi.org/10.1016/j.mimet.2018.10.004
        • Idelevich E.A.
        • Becker K.
        How to accelerate antimicrobial susceptibility testing.
        Clin Microbiol Infect. 2019; 25: 1347-1355https://doi.org/10.1016/j.cmi.2019.04.025
        • Hazelton B.
        • Thomas L.C.
        • Olma T.
        • Kok J.
        • O'Sullivan M.
        • Chen S.C.-A.
        • et al.
        Rapid and accurate direct antibiotic susceptibility testing of blood culture broths using MALDI Sepsityper combined with the BD Phoenix automated system.
        J Med Microbiol. 2014; 63: 1590-1594https://doi.org/10.1099/jmm.0.075580-0
        • Cordovana M.
        • Zignoli A.
        • Ambretti S.
        Rapid Sepsityper in clinical routine: 2 years' successful experience.
        J Med Microbiol. 2020; 69: 1398-1404https://doi.org/10.1099/jmm.0.001268
        • (CLSI) CaLSI
        Performance standards for antimicrobial susceptibility testing; 22nd informational supplement (M100–S22).
        Clinical and Laboratory Standards Institute, Wayne, PA2012
        • Gütgemann F.
        • Müller A.
        • Churin Y.
        • Jung A.
        • Braun A.S.
        • Yue M.
        • et al.
        Development of a harmonized method for antimicrobial susceptibility testing of Bordetella avium using broth microdilution and detection of resistance genes.
        J Appl Microbiol. 2021; https://doi.org/10.1111/jam.15305
        • Morgenthaler N.G.
        • Kostrzewa M.
        Rapid identification of pathogens in positive blood culture of patients with sepsis: Review and meta-analysis of the performance of the sepsityper kit.
        Int J Microbiol. 2015; (2015)827416https://doi.org/10.1155/2015/827416
        • Yonetani S.
        • Ohnishi H.
        • Ohkusu K.
        • Matsumoto T.
        • Watanabe T.
        Direct identification of microorganisms from positive blood cultures by MALDI-TOF MS using an in-house saponin method.
        Int J Infect Dis. 2016; 52: 37-42https://doi.org/10.1016/j.ijid.2016.09.014
        • Kayin M.
        • Mert B.
        • Aydemir S.
        • Özenci V.
        Comparison of rapid BACpro® II, Sepsityper® kit and in-house preparation methods for direct identification of bacteria from blood cultures by MALDI-TOF MS with and without Sepsityper® module analysis.
        Eur J Clin Microbiol Infect Dis. 2019; 38: 2133-2143https://doi.org/10.1007/s10096-019-03654-4
        • Martinez R.M.
        • Bauerle E.R.
        • Fang F.C.
        • Butler-Wu S.M.
        • Patel R.
        Evaluation of three rapid diagnostic methods for direct identification of microorganisms in positive blood cultures.
        J Clin Microbiol. 2014; 52: 2521-2529https://doi.org/10.1128/JCM.00529-14
        • Höring S.
        • Massarani A.S.
        • Löffler B.
        • Rödel J.
        Rapid antibiotic susceptibility testing in blood culture diagnostics performed by direct inoculation using the VITEK®-2 and BD Phoenix™ platforms.
        Eur J Clin Microbiol Infect Dis. 2019; 38: 471-478https://doi.org/10.1007/s10096-018-03445-3
        • Beuving J.
        • van der Donk C.F.M.
        • Linssen C.F.M.
        • Wolffs P.F.G.
        • Verbon A.
        Evaluation of direct inoculation of the BD PHOENIX system from positive BACTEC blood cultures for both Gram-positive cocci and Gram-negative rods.
        BMC Microbiol. 2011; 156: 11https://doi.org/10.1186/1471-2180-11-156
        • Boland L.
        • Streel C.
        • De Wolf H.
        • Rodriguez H.
        • Verroken A.
        Rapid antimicrobial susceptibility testing on positive blood cultures through an innovative light scattering technology: performances and turnaround time evaluation.
        BMC Infect Dis. 2019; 989: 19https://doi.org/10.1186/s12879-019-4623-x