• Media type: E-Article
  • Title: Novel flow cytometric screening method for bacterial contamination of red blood cells: a proof‐of‐principle evaluation
  • Contributor: Vollmer, Tanja; Knabbe, Cornelius; Dreier, Jens
  • Published: Wiley, 2014
  • Published in: Transfusion, 54 (2014) 3pt2, Seite 900-909
  • Language: English
  • DOI: 10.1111/trf.12513
  • ISSN: 0041-1132; 1537-2995
  • Keywords: Hematology ; Immunology ; Immunology and Allergy
  • Origination:
  • Footnote:
  • Description: <jats:sec><jats:title>Background</jats:title><jats:p>Platelet concentrates (<jats:styled-content style="fixed-case">PC</jats:styled-content>s) are the major source of transfusion‐transmitted bacterial infection but the significantly higher number of transfused red blood cells (<jats:styled-content style="fixed-case">RBC</jats:styled-content>s) has resulted in a similar occurrence of possible transfusion‐associated bacterial infections as those of recipients transfused with platelets. The aim of this study was adaption of the <jats:styled-content style="fixed-case">B</jats:styled-content>acti<jats:styled-content style="fixed-case">F</jats:styled-content>low (<jats:styled-content style="fixed-case">BF</jats:styled-content>) flow cytometric rapid <jats:styled-content style="fixed-case">PC</jats:styled-content> screening method for the detection of bacterial contamination in <jats:styled-content style="fixed-case">RBC</jats:styled-content>s.</jats:p></jats:sec><jats:sec><jats:title>Study Design and Methods</jats:title><jats:p>The <jats:styled-content style="fixed-case">BF</jats:styled-content> assay was used to detect and count bacteria based on esterase activity in viable cells in <jats:styled-content style="fixed-case">RBC</jats:styled-content>s. The initial sample preparation for <jats:styled-content style="fixed-case">RBC</jats:styled-content>s included the lysis of <jats:styled-content style="fixed-case">RBCs</jats:styled-content>, followed by enrichment of bacteria by centrifugation and subsequent standard <jats:styled-content style="fixed-case">PC</jats:styled-content> sample preparation. Two‐hundred <jats:styled-content style="fixed-case">RBC</jats:styled-content> units were analyzed by <jats:styled-content style="fixed-case">BF</jats:styled-content> and the <jats:styled-content style="fixed-case">B</jats:styled-content>ac<jats:styled-content style="fixed-case">T</jats:styled-content>/<jats:styled-content style="fixed-case">A</jats:styled-content>lert automated culture system. Furthermore, <jats:styled-content style="fixed-case">RBC</jats:styled-content>s were spiked with eight different bacteria to monitor bacterial growth kinetics.</jats:p></jats:sec><jats:sec><jats:title>Results</jats:title><jats:p>The <jats:styled-content style="fixed-case">BF</jats:styled-content> showed an excellent correlation to conventional plate count results after <jats:styled-content style="fixed-case">RBC</jats:styled-content> sample processing. The diagnostic sensitivity of the assay was determined to fewer than 500 counts/<jats:styled-content style="fixed-case">mL</jats:styled-content>. Analysis of <jats:styled-content style="fixed-case">RBC</jats:styled-content> units showed concordant negative results by <jats:styled-content style="fixed-case">BF</jats:styled-content> and culture. Growth kinetics of bacteria were successfully monitored using flow cytometry, showing that <jats:italic><jats:styled-content style="fixed-case">Y</jats:styled-content>ersinia enterocolitica</jats:italic> or <jats:italic><jats:styled-content style="fixed-case">S</jats:styled-content>erratia liquefaciens</jats:italic> had the capability to grow in <jats:styled-content style="fixed-case">RBC</jats:styled-content>s to high counts.</jats:p></jats:sec><jats:sec><jats:title>Conclusion</jats:title><jats:p>Our study demonstrates the successful modification and application of a rapid flow cytometric screening method for bacterial contamination in <jats:styled-content style="fixed-case">RBC</jats:styled-content>s. The <jats:styled-content style="fixed-case">BF</jats:styled-content> assay represents a powerful basic tool to study bacterial contamination, potential screening strategies of <jats:styled-content style="fixed-case">RBC</jats:styled-content>s, and the clinical outcome of screened <jats:styled-content style="fixed-case">RBC</jats:styled-content>s prepared for transfusion.</jats:p></jats:sec>