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A highly sensitive multiplex immunoassay for inflammatory cytokines in dried blood spots
Objectives Inflammatory cytokines are key regulators of inflammation, but current measurement approaches require venous blood to quantify low circulating concentrations associated with chronic, low‐grade inflammation. This article describes a highly sensitive multiplex immunoassay protocol for the m...
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| Published in: | American journal of human biology 2021-11, Vol.33 (6), p.e23558-n/a |
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| Main Authors: | , , , , |
| Format: | Article |
| Language: | English |
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| cites | cdi_FETCH-LOGICAL-c3578-c09b93914e6d37906d18bc108fbeeac239202d173d4b7918bf340531229b3c2f3 |
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| container_issue | 6 |
| container_start_page | e23558 |
| container_title | American journal of human biology |
| container_volume | 33 |
| creator | McDade, Thomas W. Miller, Aaron Tran, Tina T. Borders, Ann E. B. Miller, Greg |
| description | Objectives
Inflammatory cytokines are key regulators of inflammation, but current measurement approaches require venous blood to quantify low circulating concentrations associated with chronic, low‐grade inflammation. This article describes a highly sensitive multiplex immunoassay protocol for the measurement of IL6, IL8, IL10, and TNFα in finger stick dried blood spot (DBS) samples.
Methods
The protocol uses a multiplex electrochemiluminescent immunoassay platform. The following measures of assay performance were evaluated: reliability (inter‐assay percent coefficient of variation; %CV), precision (intra‐assay %CV), lower limit of detection (LLD), linearity of dilution, and agreement with results from matched plasma samples.
Results
Analysis of three control samples across the assay range indicated an acceptable level of precision and reliability for each cytokine. Linearity of dilution returned average values that ranged from 104.1 to 127.6% of expected. Lower limits of detection for IL6, IL8, and IL10 were |
| doi_str_mv | 10.1002/ajhb.23558 |
| format | article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_2474465463</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2474465463</sourcerecordid><originalsourceid>FETCH-LOGICAL-c3578-c09b93914e6d37906d18bc108fbeeac239202d173d4b7918bf340531229b3c2f3</originalsourceid><addsrcrecordid>eNp9kE1P3DAQhi1UxMLCpT-gstRLhZRl_JXExwVBKVqJC0jcLCdxul6ceLGTlvx7TJdy4MBpRjOPXs08CH0lsCAA9Exv1tWCMiHKPXRIBIUsZwBfUg-cZiAYm6GjGDcAIHMoD9CMMVZSkueH6GGJ1_b32k04mj7awf4xuBvdYLfOPGPbdWPvdYx6wq0P2Pat012nBx8mXE-Df7S9iWmMm2BNgyvnfYPj1g_xGO232kVz8lbn6P7q8u7iOlvd_vx1sVxlNRNFmdUgK8kk4SZvWCEhb0hZ1QTKtjJG15RJCrQhBWt4Vci0axlPHxFKZcVq2rI5-rHL3Qb_NJo4qM7G2jine-PHqCgvOM8Fz1lCv39AN34MfbpOUSEFF5yLIlGnO6oOPsZgWrUNttNhUgTUq2_16lv9853gb2-RY9WZ5h39LzgBZAf8tc5Mn0Sp5c31-S70BYrlijY</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2595454457</pqid></control><display><type>article</type><title>A highly sensitive multiplex immunoassay for inflammatory cytokines in dried blood spots</title><source>Wiley</source><creator>McDade, Thomas W. ; Miller, Aaron ; Tran, Tina T. ; Borders, Ann E. B. ; Miller, Greg</creator><creatorcontrib>McDade, Thomas W. ; Miller, Aaron ; Tran, Tina T. ; Borders, Ann E. B. ; Miller, Greg</creatorcontrib><description>Objectives
Inflammatory cytokines are key regulators of inflammation, but current measurement approaches require venous blood to quantify low circulating concentrations associated with chronic, low‐grade inflammation. This article describes a highly sensitive multiplex immunoassay protocol for the measurement of IL6, IL8, IL10, and TNFα in finger stick dried blood spot (DBS) samples.
Methods
The protocol uses a multiplex electrochemiluminescent immunoassay platform. The following measures of assay performance were evaluated: reliability (inter‐assay percent coefficient of variation; %CV), precision (intra‐assay %CV), lower limit of detection (LLD), linearity of dilution, and agreement with results from matched plasma samples.
Results
Analysis of three control samples across the assay range indicated an acceptable level of precision and reliability for each cytokine. Linearity of dilution returned average values that ranged from 104.1 to 127.6% of expected. Lower limits of detection for IL6, IL8, and IL10 were <0.5, and <1.0 pg/ml for TNFα. Level of agreement in results between matched DBS and plasma samples was high for all cytokines except for IL8.
Conclusions
Finger stick DBS sampling provides a viable alternative to venipuncture for the quantification of IL6, IL10, and TNFα at low concentrations associated with chronic inflammation. The presence of red blood cells may interfere with the quantification of IL8 in DBS. In facilitating blood collection in nonclinical settings this method can advance scientific understandings of how social and ecological contexts shape immune function and health over the life course.</description><identifier>ISSN: 1042-0533</identifier><identifier>EISSN: 1520-6300</identifier><identifier>DOI: 10.1002/ajhb.23558</identifier><identifier>PMID: 33382166</identifier><language>eng</language><publisher>Hoboken, USA: John Wiley & Sons, Inc</publisher><subject>Blood circulation ; Coefficient of variation ; Cytokines ; Cytokines - analysis ; Dilution ; Dried Blood Spot Testing ; Electrochemiluminescence ; Erythrocytes ; Humans ; Immune response ; Immunoassay ; Inflammation ; Interleukin 1 ; Interleukin 10 ; Interleukin 6 ; Interleukin 8 ; Linearity ; Low concentrations ; Multiplexing ; Reliability analysis ; Reproducibility of Results ; Tumor necrosis factor-α</subject><ispartof>American journal of human biology, 2021-11, Vol.33 (6), p.e23558-n/a</ispartof><rights>2020 Wiley Periodicals LLC.</rights><rights>2021 Wiley Periodicals LLC.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3578-c09b93914e6d37906d18bc108fbeeac239202d173d4b7918bf340531229b3c2f3</citedby><cites>FETCH-LOGICAL-c3578-c09b93914e6d37906d18bc108fbeeac239202d173d4b7918bf340531229b3c2f3</cites><orcidid>0000-0003-2744-8916 ; 0000-0001-5829-648X</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27903,27904</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/33382166$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>McDade, Thomas W.</creatorcontrib><creatorcontrib>Miller, Aaron</creatorcontrib><creatorcontrib>Tran, Tina T.</creatorcontrib><creatorcontrib>Borders, Ann E. B.</creatorcontrib><creatorcontrib>Miller, Greg</creatorcontrib><title>A highly sensitive multiplex immunoassay for inflammatory cytokines in dried blood spots</title><title>American journal of human biology</title><addtitle>Am J Hum Biol</addtitle><description>Objectives
Inflammatory cytokines are key regulators of inflammation, but current measurement approaches require venous blood to quantify low circulating concentrations associated with chronic, low‐grade inflammation. This article describes a highly sensitive multiplex immunoassay protocol for the measurement of IL6, IL8, IL10, and TNFα in finger stick dried blood spot (DBS) samples.
Methods
The protocol uses a multiplex electrochemiluminescent immunoassay platform. The following measures of assay performance were evaluated: reliability (inter‐assay percent coefficient of variation; %CV), precision (intra‐assay %CV), lower limit of detection (LLD), linearity of dilution, and agreement with results from matched plasma samples.
Results
Analysis of three control samples across the assay range indicated an acceptable level of precision and reliability for each cytokine. Linearity of dilution returned average values that ranged from 104.1 to 127.6% of expected. Lower limits of detection for IL6, IL8, and IL10 were <0.5, and <1.0 pg/ml for TNFα. Level of agreement in results between matched DBS and plasma samples was high for all cytokines except for IL8.
Conclusions
Finger stick DBS sampling provides a viable alternative to venipuncture for the quantification of IL6, IL10, and TNFα at low concentrations associated with chronic inflammation. The presence of red blood cells may interfere with the quantification of IL8 in DBS. In facilitating blood collection in nonclinical settings this method can advance scientific understandings of how social and ecological contexts shape immune function and health over the life course.</description><subject>Blood circulation</subject><subject>Coefficient of variation</subject><subject>Cytokines</subject><subject>Cytokines - analysis</subject><subject>Dilution</subject><subject>Dried Blood Spot Testing</subject><subject>Electrochemiluminescence</subject><subject>Erythrocytes</subject><subject>Humans</subject><subject>Immune response</subject><subject>Immunoassay</subject><subject>Inflammation</subject><subject>Interleukin 1</subject><subject>Interleukin 10</subject><subject>Interleukin 6</subject><subject>Interleukin 8</subject><subject>Linearity</subject><subject>Low concentrations</subject><subject>Multiplexing</subject><subject>Reliability analysis</subject><subject>Reproducibility of Results</subject><subject>Tumor necrosis factor-α</subject><issn>1042-0533</issn><issn>1520-6300</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><recordid>eNp9kE1P3DAQhi1UxMLCpT-gstRLhZRl_JXExwVBKVqJC0jcLCdxul6ceLGTlvx7TJdy4MBpRjOPXs08CH0lsCAA9Exv1tWCMiHKPXRIBIUsZwBfUg-cZiAYm6GjGDcAIHMoD9CMMVZSkueH6GGJ1_b32k04mj7awf4xuBvdYLfOPGPbdWPvdYx6wq0P2Pat012nBx8mXE-Df7S9iWmMm2BNgyvnfYPj1g_xGO232kVz8lbn6P7q8u7iOlvd_vx1sVxlNRNFmdUgK8kk4SZvWCEhb0hZ1QTKtjJG15RJCrQhBWt4Vci0axlPHxFKZcVq2rI5-rHL3Qb_NJo4qM7G2jine-PHqCgvOM8Fz1lCv39AN34MfbpOUSEFF5yLIlGnO6oOPsZgWrUNttNhUgTUq2_16lv9853gb2-RY9WZ5h39LzgBZAf8tc5Mn0Sp5c31-S70BYrlijY</recordid><startdate>202111</startdate><enddate>202111</enddate><creator>McDade, Thomas W.</creator><creator>Miller, Aaron</creator><creator>Tran, Tina T.</creator><creator>Borders, Ann E. B.</creator><creator>Miller, Greg</creator><general>John Wiley & Sons, Inc</general><general>Wiley Subscription Services, Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QP</scope><scope>7SN</scope><scope>7ST</scope><scope>7T5</scope><scope>7TK</scope><scope>7TM</scope><scope>7TS</scope><scope>C1K</scope><scope>H94</scope><scope>K9.</scope><scope>SOI</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0003-2744-8916</orcidid><orcidid>https://orcid.org/0000-0001-5829-648X</orcidid></search><sort><creationdate>202111</creationdate><title>A highly sensitive multiplex immunoassay for inflammatory cytokines in dried blood spots</title><author>McDade, Thomas W. ; Miller, Aaron ; Tran, Tina T. ; Borders, Ann E. B. ; Miller, Greg</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3578-c09b93914e6d37906d18bc108fbeeac239202d173d4b7918bf340531229b3c2f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>Blood circulation</topic><topic>Coefficient of variation</topic><topic>Cytokines</topic><topic>Cytokines - analysis</topic><topic>Dilution</topic><topic>Dried Blood Spot Testing</topic><topic>Electrochemiluminescence</topic><topic>Erythrocytes</topic><topic>Humans</topic><topic>Immune response</topic><topic>Immunoassay</topic><topic>Inflammation</topic><topic>Interleukin 1</topic><topic>Interleukin 10</topic><topic>Interleukin 6</topic><topic>Interleukin 8</topic><topic>Linearity</topic><topic>Low concentrations</topic><topic>Multiplexing</topic><topic>Reliability analysis</topic><topic>Reproducibility of Results</topic><topic>Tumor necrosis factor-α</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>McDade, Thomas W.</creatorcontrib><creatorcontrib>Miller, Aaron</creatorcontrib><creatorcontrib>Tran, Tina T.</creatorcontrib><creatorcontrib>Borders, Ann E. B.</creatorcontrib><creatorcontrib>Miller, Greg</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Ecology Abstracts</collection><collection>Environment Abstracts</collection><collection>Immunology Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Physical Education Index</collection><collection>Environmental Sciences and Pollution Management</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Environment Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>American journal of human biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>McDade, Thomas W.</au><au>Miller, Aaron</au><au>Tran, Tina T.</au><au>Borders, Ann E. B.</au><au>Miller, Greg</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A highly sensitive multiplex immunoassay for inflammatory cytokines in dried blood spots</atitle><jtitle>American journal of human biology</jtitle><addtitle>Am J Hum Biol</addtitle><date>2021-11</date><risdate>2021</risdate><volume>33</volume><issue>6</issue><spage>e23558</spage><epage>n/a</epage><pages>e23558-n/a</pages><issn>1042-0533</issn><eissn>1520-6300</eissn><abstract>Objectives
Inflammatory cytokines are key regulators of inflammation, but current measurement approaches require venous blood to quantify low circulating concentrations associated with chronic, low‐grade inflammation. This article describes a highly sensitive multiplex immunoassay protocol for the measurement of IL6, IL8, IL10, and TNFα in finger stick dried blood spot (DBS) samples.
Methods
The protocol uses a multiplex electrochemiluminescent immunoassay platform. The following measures of assay performance were evaluated: reliability (inter‐assay percent coefficient of variation; %CV), precision (intra‐assay %CV), lower limit of detection (LLD), linearity of dilution, and agreement with results from matched plasma samples.
Results
Analysis of three control samples across the assay range indicated an acceptable level of precision and reliability for each cytokine. Linearity of dilution returned average values that ranged from 104.1 to 127.6% of expected. Lower limits of detection for IL6, IL8, and IL10 were <0.5, and <1.0 pg/ml for TNFα. Level of agreement in results between matched DBS and plasma samples was high for all cytokines except for IL8.
Conclusions
Finger stick DBS sampling provides a viable alternative to venipuncture for the quantification of IL6, IL10, and TNFα at low concentrations associated with chronic inflammation. The presence of red blood cells may interfere with the quantification of IL8 in DBS. In facilitating blood collection in nonclinical settings this method can advance scientific understandings of how social and ecological contexts shape immune function and health over the life course.</abstract><cop>Hoboken, USA</cop><pub>John Wiley & Sons, Inc</pub><pmid>33382166</pmid><doi>10.1002/ajhb.23558</doi><tpages>7</tpages><orcidid>https://orcid.org/0000-0003-2744-8916</orcidid><orcidid>https://orcid.org/0000-0001-5829-648X</orcidid></addata></record> |
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| source | Wiley |
| subjects | Blood circulation Coefficient of variation Cytokines Cytokines - analysis Dilution Dried Blood Spot Testing Electrochemiluminescence Erythrocytes Humans Immune response Immunoassay Inflammation Interleukin 1 Interleukin 10 Interleukin 6 Interleukin 8 Linearity Low concentrations Multiplexing Reliability analysis Reproducibility of Results Tumor necrosis factor-α |
| title | A highly sensitive multiplex immunoassay for inflammatory cytokines in dried blood spots |
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