Journals / Türk Biyokimya Dergisi / 2018 / Cilt: 43 - Sayı: 5

Comparison of ELISA and flow cytometry for measurement of interleukin-1 beta, interleukin-6 and tumor necrosis factor-α

Pages
540–548
Publication Date
2022-11-05
DOI
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AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

However, our study hasa small number of participants, hence this study need to beconfirmed by investigations involving more participants.

📌 3 Key Takeaways

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Objective & Hypothesis:Background: Although majority of the previous studieshave shown a good correlation between enzyme linkedimmuno sorbent assay (ELISA) and flow cytometry interms of cytokines, two laboratory methods usually werecompared with the regression analysis and correlation inthe literature.
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Key Findings & Data:This study aimed at comparing the ELISAand flow cytometry assay for measuring cytokines byusing two different statistical methods, regression analysisand Bland-Altman plot.Materials and methods: Fifty patients, diagnosed withhypercholesterolemia and expecting high level serumcytokines, and 30 healthy volunteers, expecting normallevel serum cytokines, were enrolled in the study.
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Conclusion & Impact:However, our study hasa small number of participants, hence this study need to beconfirmed by investigations involving more participants.
🔬Design:Research Article
📍Scope:Comprehensive Sample
⚙️Variables:Domain Parameters

Abstract

Background: Although majority of the previous studieshave shown a good correlation between enzyme linkedimmuno sorbent assay (ELISA) and flow cytometry interms of cytokines, two laboratory methods usually werecompared with the regression analysis and correlation inthe literature. This study aimed at comparing the ELISAand flow cytometry assay for measuring cytokines byusing two different statistical methods, regression analysisand Bland-Altman plot.Materials and methods: Fifty patients, diagnosed withhypercholesterolemia and expecting high level serumcytokines, and 30 healthy volunteers, expecting normallevel serum cytokines, were enrolled in the study. The interleukin-1 beta (IL-1β), interleukin-6 (IL-6) and tumor necrosisfactor-alpha (TNF-α) were measured using ELISA, and comparedwith obtained levels using flow cytometric method.Results: Although regression analysis showed that the twomethods are compatible with measurements of IL-1β, IL-6and TNF-α, they tended to show dissimilarity with measurementsof IL-1β and TNF-α based on Bland-Altman graphs.Conclusion: According to Bland-Altman plot, our resultsproviding evidence of ELISA and flow cytometry assays werecompatible with each other for IL-1β and IL-6 measurementscompared to TNF-α measurement. However, our study hasa small number of participants, hence this study need to beconfirmed by investigations involving more participants.

Özet

Amaç: Önceki çalışmaların büyük çoğunluğu, sitokinler açısından Enzim Bağlı İmmüno Sorbent Testi (ELISA) ile akış sitometrisi arasında iyi bir korelasyon olduğunu göstermesine rağmen, iki laboratuvar yöntemi literatürde genellikle regresyon analizi ve korelasyon ile karşılaştırılmıştır. Bu çalışmada, iki farklı istatistiksel yöntem olan regresyon analizi ve Bland-Altman grafiği kullanılarak sitokinlerin ölçümü için ELISA ve akış sitometri analizinin karşılaştırılması amaçlanmıştır. Gereç ve Yöntem: Hiperkolesterolemi tanısı konulan ve yüksek düzeyde serum sitokin düzeyleri beklenen elli hasta ve normal serum sitokin düzeyleri beklenen 30 sağlıklı gönüllü çalışmaya dâhil edildi. İnterlökin-1 beta (IL-1β), interlökin-6 (IL-6) ve tümör nekrozis faktör-alfa (TNF-α) ELISA kullanılarak ölçüldü ve akış sitometri yöntemi kullanılarak elde edilen düzeylerle karşılaştırıldı. Bulgular: Regresyon analizi, iki yöntemin IL-1β, IL-6 ve TNF-α’nın ölçülmesinde uyumlu olduklarını göstermesine rağmen, Bland-Altman grafiğine göre iki yöntemin IL-1β ve TNF-α’nın ölçümünde farklılık gösterme eğiliminde olduğu belirlendi. Sonuç: Bland-Altman grafiğine göre; sonuçlarımız, ELISA ve akış sitometrisi analizlerinin IL-1β ve IL-6 ölçümünde, TNF-α ölçümüne göre daha uyumlu olduğunu kanıtlamaktadır. Bununla birlikte, çalışmamızın az sayıda katılımcı içermesi nedeniyle daha fazla katılımcıyı içeren araştırmalarla bu çalışmanın doğrulanması gerekmektedir.

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Cite This Article

CİMEN, L., ÇETİN, İ., KOKER, M. Y., ÖZTÜRK, A., ÇİMEN, B., ÇETİN, A., ŞEN, A., SAVAŞ, G. (2022). Comparison of ELISA and flow cytometry for measurement of interleukin-1 beta, interleukin-6 and tumor necrosis factor-α. Türk Biyokimya Dergisi, Cilt: 43 - Sayı: 5, 540–548. https://ajindex.com/articles/620933

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References (15)

  1. Chaudhry H, Zhou J, Zhong Y, Ali MM, McGuire F, Nagarkatti PS, et al. Role of cytokines as a double-edged sword in sepsis. In Vivo 2013;27:669–84
  2. Elsabahy M, Wooley KL. Cytokines as biomarkers of nanoparticle immunotoxicity. Chem Soc Rev 2013;42:5552–76
  3. Skogstrand K. Multiplex assays of inflammatory markers, a description of methods and discussion of precautions – our experience through the last ten years. Methods 2012;56:204–12
  4. Leng SX, McElhaney JE, Walston JD, Xie D, Fedarko NS, Kuchel GA. ELISA and multiplex technologies for cytokine measurement in inflammation and aging research. J Gerontol A Biol Sci Med Sci 2008;63:879–84
  5. Freer G, Rindi L. Intracellular cytokine detection by fluorescenceactivated flow cytometry: basic principles and recent advances. Methods 2013;61:30–8
  6. Daubeuf F, Becker J, Aguilar-Pimentel JA, Ebel C, Hrabě de Angelis M, Hérault Y, et al. A fast, easy, and customizable eight-color flow cytometric method for analysis of the cellular content of bronchoalveolar lavage fluid in the mouse. Curr Protoc Mouse Biol 2017;7:88–99
  7. Dabitao D, Margolick JB, Lopez J, Bream JH. Multiplex measurement of proinflammatory cytokines in human serum: comparison of the meso scale discovery electrochemiluminescence assay and the cytometric bead array. J Immunol Methods 2011;372:71–7
  8. Stenken JA, Poschenrieder AJ. Bioanalytical chemistry of cytokines – a review. Anal Chim Acta 2015;853:95–115
  9. Rountree W, Vandergrift N, Bainbridge J, Sanchez AM, Denny TN. Statistical methods for the assessment of EQAPOL proficiency testing: ELISpot, luminex, and flow cytometry. J Immunol Methods 2014;409:72–81
  10. Giavarina D. Understanding Bland Altman analysis. Biochem Med 2015;25:141–51
  11. Scott LE, Galpin JS, Glencross DK. Multiple method comparison: statistical model using percentage similarity. Cytometry B Clin Cytom 2003;54:46–53
  12. Carson RT, Vignali DA. Simultaneous quantitation of 15 cytokines using a multiplexed flow cytometric assay. J Immunol Methods 1999;227:41–52
  13. Chen R, Lowe L, Wilson JD, Crowther E, Tzeggai K, Bishop JE, et al. Simultaneous quantification of six human cytokines in a single sample using microparticle-based flow cytometric technology. Clin Chem 1999;45:1693–4
  14. Kellar KL, Kalwar RR, Dubois KA, Crouse D, Chafin WD, Kane BE. Multiplexed fluorescent bead-based immunoassays for quantitation of human cytokines in serum and culture supernatants. Cytometry 2001;45:27–36
  15. Hanneman SK. Design, analysi