BACKGROUND: Biomarker assays are often conducted on whole blood samples in the course of drug development studies. Because bacterial lipopolysaccharide (LPS) (endotoxin) contamination is known to cause spontaneous cytokine production by monocytes, contamination of blood collection tubes may interfere with biomarker assay results. METHODS: Whole blood from healthy donors was collected into plastic or glass sodium (Na(+))-heparin Vacutainer() blood collection tubes and heparinized syringes. Samples were analyzed for phosphoprotein response, cytokine production, and RNA expression. Tubes were tested for endotoxin contamination by use of the limulus amoebocyte lysate assay. RESULTS: Results of phospho-flow cytometry, branched DNA (bDNA), and ELISA assays indicated that a specific lot (#5339582) of plastic Na(+)-heparin Vacutainer tubes was highly contaminated with an endotoxinlike substance, and contamination was confirmed by the limulus amoebocyte lysate assay. Analysis of multiple-analyte panels revealed that analytes whose changed expression was predictive of LPS stimulation were increased when whole blood was incubated in contaminated tubes for 6 or 18 h. Two additional lots of plastic tubes tested had detectable amounts of endotoxin sufficient to strongly alter phospho-flow cytometry analyses, as determined by the fold change in phosphorylation of p38 mitogen-activated protein kinase in response to tumor necrosis factor alpha and LPS. In contrast, 3 lots of glass tubes had substantially lower levels of spontaneous blood activation. CONCLUSIONS: Endotoxin contamination associated with tubes from 3 lots of a particular type of plastic Na(+)-heparin Vacutainer tube dramatically affected biomarker assay measurements. Prescreening these tubes is suggested before their use in clinical sample analysis.
BACKGROUND: Biomarker assays are often conducted on whole blood samples in the course of drug development studies. Because bacterial lipopolysaccharide (LPS) (endotoxin) contamination is known to cause spontaneous cytokine production by monocytes, contamination of blood collection tubes may interfere with biomarker assay results. METHODS: Whole blood from healthy donors was collected into plastic or glass sodium (Na(+))-heparin Vacutainer() blood collection tubes and heparinized syringes. Samples were analyzed for phosphoprotein response, cytokine production, and RNA expression. Tubes were tested for endotoxin contamination by use of the limulus amoebocyte lysate assay. RESULTS: Results of phospho-flow cytometry, branched DNA (bDNA), and ELISA assays indicated that a specific lot (#5339582) of plastic Na(+)-heparin Vacutainer tubes was highly contaminated with an endotoxinlike substance, and contamination was confirmed by the limulus amoebocyte lysate assay. Analysis of multiple-analyte panels revealed that analytes whose changed expression was predictive of LPS stimulation were increased when whole blood was incubated in contaminated tubes for 6 or 18 h. Two additional lots of plastic tubes tested had detectable amounts of endotoxin sufficient to strongly alter phospho-flow cytometry analyses, as determined by the fold change in phosphorylation of p38 mitogen-activated protein kinase in response to tumor necrosis factor alpha and LPS. In contrast, 3 lots of glass tubes had substantially lower levels of spontaneous blood activation. CONCLUSIONS: Endotoxin contamination associated with tubes from 3 lots of a particular type of plastic Na(+)-heparin Vacutainer tube dramatically affected biomarker assay measurements. Prescreening these tubes is suggested before their use in clinical sample analysis.
Authors: T N Nissen; N M Birk; B A Blok; R J W Arts; A Andersen; J Kjærgaard; L M Thøstesen; T Hoffmann; D L Jeppesen; S D Nielsen; P-E Kofoed; L G Stensballe; P Aaby; M Ruhwald; M G Netea; C S Benn; O Pryds Journal: Eur J Clin Microbiol Infect Dis Date: 2017-09-10 Impact factor: 3.267
Authors: Juan C Nieto; Elisabet Cantó; Carlos Zamora; M Angels Ortiz; Cándido Juárez; Silvia Vidal Journal: PLoS One Date: 2012-03-05 Impact factor: 3.240
Authors: Krista Kuuliala; Antti Kuuliala; Riitta Koivuniemi; Suvi Oksanen; Mari Hämäläinen; Eeva Moilanen; Hannu Kautiainen; Marjatta Leirisalo-Repo; Heikki Repo Journal: PLoS One Date: 2015-09-09 Impact factor: 3.240
Authors: Fred K Tabung; Brenda M Birmann; Mara M Epstein; Otoniel Martínez-Maza; Elizabeth C Breen; Kana Wu; Edward L Giovannucci Journal: Curr Dev Nutr Date: 2017-10-17
Authors: Ingrid Elisia; Vivian Lam; Brandon Cho; Mariah Hay; Michael Yu Li; Jordanna Kapeluto; Tom Elliott; David Harris; Luke Bu; William Jia; Hilary Leung; William Mohn; Gerald Krystal Journal: PLoS One Date: 2020-02-06 Impact factor: 3.240