Multiplex analysis of cytokines in exhaled breath condensate

Sack, Ulrich; Scheibe, R.; Wötzel, Michael; Hammerschmidt, Stefan; Kühn, Hartmut; Emmrich, Frank; Hoheisel, Gerard; Wirtz, Hubert; Geßner, Christian

doi:10.1002/cyto.a.20231

Cited by 90 publications

(86 citation statements)

References 12 publications

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“…The overall level of detection was 46-97%, which was much better compared with cytometric bead array in small samples in children (,50%), and in large 1,000-2,000-mL lyophilisated samples in adults (3-100%) [6,[23][24][25][26]. When compared with conventional (solid-phase) immunoassays, multiplexed immunoassays detect bioactive and inactive molecules, have a growing analytical range, are rapid (take hours instead of days to perform), have good precision (CV 10-15%), are not interfered with by drugs, and have simple protocols [7].…”

Section: Discussionmentioning

confidence: 95%

Biomarker reproducibility in exhaled breath condensate collected with different condensers

Rosias¹,

Robroeks²,

Kester³

et al. 2008

Eur Respir J

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Optimal collection and analysis of exhaled breath condensate (EBC) are prerequisites for standardisation and reproducibility of assessments. The present study aimed to assess reproducibility of EBC volume, hydrogen peroxide (H 2 O 2 ), 8-isoprostane and cytokine measurements using different condensers, including a newly developed glass condenser.At four points in time, 30 healthy subjects performed sequential EBC collections randomly using the following four condensers: glass, silicone, EcoScreen1 (Erich Jaeger GmbH, Hoechberg, Germany) and an optimised glass condenser. In small EBC samples, H 2 O 2 was measured by spectrophotometer, 8-isoprostane by enzyme immunoassay, and cytokines by multiplexed xMAP1 technology (Luminex Corporation, Austin, TX, USA).The optimised glass condenser yielded significantly more EBC volume (median 2,025 mL, interquartile range 1,600-2,525). The reproducibility of EBC volume, yielded by the new glass condenser, was comparable with EcoScreen1 (19-20 coefficients of variation (CV)%), but was significantly better compared with silicone and glass (29-37 CV%). The new condenser was associated with significantly more detections of H 2 O 2 , 8-isoprostane, interleukin-2, -4, -5 and -13, and tumour necrosis factor-a. Isoprostane concentrations were significantly higher using the new condenser, whereas H 2 O 2 and cytokine concentrations were not. Reproducibility of biomarkers was equally variable for all condenser types.In conclusion, significantly more exhaled breath condensate volume and biomarker detections were found using the optimised glass condenser, including higher 8-isoprostane levels. However, biomarker reproducibility in exhaled breath condensate in healthy adults was not influenced by the type of condenser.

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Section: Discussionmentioning

confidence: 95%

Biomarker reproducibility in exhaled breath condensate collected with different condensers

Rosias¹,

Robroeks²,

Kester³

et al. 2008

Eur Respir J

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“…This may be overcome by lyophilisation, which enables removal water and other volatile substances, but may degrade biomarkers [ 53 ]. EBC lyophilisation can concentrate samples up to a factor of 30 times, thus improving sensitivity [ 61 ]. There was no relationship between EBC cytokine concentrations and storage times up to 1 year in samples stored at − 80 ° C [ 157 ].…”

Section: Cytokines Chemokines and Growth Factorsmentioning

confidence: 99%

Exhaled breath condensate: a comprehensive update

Ahmadzai

Huang²,

Hettiarachchi³

et al. 2013

Clinical Chemistry and Laboratory Medicine

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Since the late 1990s, a surge in interest in the analysis of exhaled breath condensate (EBC) resulted in the American Thoracic Society and European Respiratory Society (ATS/ERS) organising a Task Force in 2001 to develop guidelines on EBC collection and measurement of biomarkers. This Task Force published their guidelines in 2005 based on literature and expert opinions at that time, and multiple shortcomings and knowledge deficits were also identified. The clinical application of EBC collection and its biomarkers are currently still limited by several of these knowledge gaps, hence further guidelines for standardisation are required to ensure external validity. Using related articles produced since the publication of the ATS/ERS Task Force report, this paper attempts to provide a comprehensive update to the original guideline and review the methodological shortcomings identified. This review can hopefully serve as a yardstick for future studies involving this emerging clinical tool.

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“…Analysis of EBC has used a variety of sensitive methods, including ELISA [1], multiplex ELISAs [1] and, more recently, mass spectroscopy [2].…”

mentioning

confidence: 99%

Validation of assays for inflammatory mediators in exhaled breath condensate

Bayley¹,

Abusriwil²,

Ahmad³

et al. 2008

Eur Respir J

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The use of exhaled breath condensate (EBC) as a tool for noninvasive assessment of lung inflammation is becoming commonplace. Many authors use commercial ELISA kits to measure inflammatory mediators within EBC. However, the very low concentrations of mediators within EBC are often below the commercially validated concentration range of the relevant ELISA and crucially below the linear part of the sigmoid standard curve. The present study seeks to validate a series of assays for use in EBC and to compare the results in EBC with those from matched sol phase sputum samples.The following mediators were measured by ELISA: leukotriene (LT)B 4 , interleukin (IL)-8, secretory leukoprotease inhibitor and a 1 -antitrypsin (AAT). Myeloperoxidase was measured by chromogenic substrate assay.Mediator concentrations reached the lower limit of quantification in only one assay (AAT) in 19.6% of subjects, while mediator concentrations reached the lower limit of detection in three assays (LTB 4 , IL-8 and AAT in 31, 6.5 and 61% of subjects, respectively). No significant correlations were present between any mediators in EBC and sol phase sputum.The results of the present study indicate that care must be exercised when interpreting mediator measurements in exhaled breath condensate and that assays must be validated at concentrations relevant to those found within the biological fluid.

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Multiplex analysis of cytokines in exhaled breath condensate

Cited by 90 publications

References 12 publications

Biomarker reproducibility in exhaled breath condensate collected with different condensers

Biomarker reproducibility in exhaled breath condensate collected with different condensers

Exhaled breath condensate: a comprehensive update

Validation of assays for inflammatory mediators in exhaled breath condensate

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