MicroRNA in vitro diagnostics using immunoassay analyzers
The implementation of new biomarkers into clinical practice is one of the most important areas in medical research. Besides their clinical impact, novel in vitro diagnostic markers promise to have a substantial effect on healthcare costs. Although numerous publications report the discovery of biomar...
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| Veröffentlicht in: | Clinical chemistry (Baltimore, Md.) Md.), 2015-04, Vol.61 (4), p.600-607 |
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| creator | Kappel, Andreas Backes, Christina Huang, Yiwei Zafari, Sachli Leidinger, Petra Meder, Benjamin Schwarz, Herbert Gumbrecht, Walter Meese, Eckart Staehler, Cord F Keller, Andreas |
| description | The implementation of new biomarkers into clinical practice is one of the most important areas in medical research. Besides their clinical impact, novel in vitro diagnostic markers promise to have a substantial effect on healthcare costs. Although numerous publications report the discovery of biomarkers, only a fraction of those markers are routinely used. One key challenge is a measurement system that is compatible with clinical workflows.
We designed a new immunoassay for microRNA (miRNA) quantification. The assay combines streptavidin-linked microparticles, a biotinylated catcher oligonucleotide complementary to a single miRNA species, and finally, a monoclonal antibody to DNA/RNA heterohybrids labeled with acridinium ester. Importantly, our assay runs on standard immunoassay analyzers. After a technical validation of the assay, we evaluated the clinical performance on 4 Alzheimer disease miRNAs.
Our assay has an analytical specificity of 99.4% and is at the same time sensitive (concentrations in the range of 1 pmol/L miRNA can be reliably profiled). Because the novel approach did not require amplification steps, we obtained high reproducibility for up to 40 biological replicates. Importantly, our assay prototype exhibited a time to result of 0.994 with the gold standard qRT-PCR.
Our miRNA immunoassay allowed the measurement of miRNA signatures with sufficient analytical sensitivity and high specificity on commonly available laboratory equipment. |
| doi_str_mv | 10.1373/clinchem.2014.232165 |
| format | Article |
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We designed a new immunoassay for microRNA (miRNA) quantification. The assay combines streptavidin-linked microparticles, a biotinylated catcher oligonucleotide complementary to a single miRNA species, and finally, a monoclonal antibody to DNA/RNA heterohybrids labeled with acridinium ester. Importantly, our assay runs on standard immunoassay analyzers. After a technical validation of the assay, we evaluated the clinical performance on 4 Alzheimer disease miRNAs.
Our assay has an analytical specificity of 99.4% and is at the same time sensitive (concentrations in the range of 1 pmol/L miRNA can be reliably profiled). Because the novel approach did not require amplification steps, we obtained high reproducibility for up to 40 biological replicates. Importantly, our assay prototype exhibited a time to result of <3 h. With human blood samples, the assay was able to measure 4 miRNAs that can detect Alzheimer disease with a diagnostic accuracy of 82% and showed a Pearson correlation >0.994 with the gold standard qRT-PCR.
Our miRNA immunoassay allowed the measurement of miRNA signatures with sufficient analytical sensitivity and high specificity on commonly available laboratory equipment.</description><identifier>ISSN: 0009-9147</identifier><identifier>EISSN: 1530-8561</identifier><identifier>DOI: 10.1373/clinchem.2014.232165</identifier><identifier>PMID: 25617425</identifier><language>eng</language><publisher>England: Oxford University Press</publisher><subject>Alzheimer Disease - blood ; Alzheimer Disease - genetics ; Alzheimer's disease ; Antibodies, Monoclonal ; Biomarkers ; Biomarkers - blood ; Blood banks ; Deoxyribonucleic acid ; Disease ; DNA ; Humans ; Immunoassay ; Immunoassay - instrumentation ; Immunoassay - methods ; Laboratories ; Medical research ; Methods ; MicroRNAs - blood ; Reproducibility of Results ; Sensitivity and Specificity</subject><ispartof>Clinical chemistry (Baltimore, Md.), 2015-04, Vol.61 (4), p.600-607</ispartof><rights>2014 American Association for Clinical Chemistry.</rights><rights>Copyright American Association for Clinical Chemistry Apr 2015</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c480t-5048c79901890c10e57c5eea8ca6784f317a45ede262e81da68da6ffc1d998f53</citedby><cites>FETCH-LOGICAL-c480t-5048c79901890c10e57c5eea8ca6784f317a45ede262e81da68da6ffc1d998f53</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25617425$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kappel, Andreas</creatorcontrib><creatorcontrib>Backes, Christina</creatorcontrib><creatorcontrib>Huang, Yiwei</creatorcontrib><creatorcontrib>Zafari, Sachli</creatorcontrib><creatorcontrib>Leidinger, Petra</creatorcontrib><creatorcontrib>Meder, Benjamin</creatorcontrib><creatorcontrib>Schwarz, Herbert</creatorcontrib><creatorcontrib>Gumbrecht, Walter</creatorcontrib><creatorcontrib>Meese, Eckart</creatorcontrib><creatorcontrib>Staehler, Cord F</creatorcontrib><creatorcontrib>Keller, Andreas</creatorcontrib><title>MicroRNA in vitro diagnostics using immunoassay analyzers</title><title>Clinical chemistry (Baltimore, Md.)</title><addtitle>Clin Chem</addtitle><description>The implementation of new biomarkers into clinical practice is one of the most important areas in medical research. Besides their clinical impact, novel in vitro diagnostic markers promise to have a substantial effect on healthcare costs. Although numerous publications report the discovery of biomarkers, only a fraction of those markers are routinely used. One key challenge is a measurement system that is compatible with clinical workflows.
We designed a new immunoassay for microRNA (miRNA) quantification. The assay combines streptavidin-linked microparticles, a biotinylated catcher oligonucleotide complementary to a single miRNA species, and finally, a monoclonal antibody to DNA/RNA heterohybrids labeled with acridinium ester. Importantly, our assay runs on standard immunoassay analyzers. After a technical validation of the assay, we evaluated the clinical performance on 4 Alzheimer disease miRNAs.
Our assay has an analytical specificity of 99.4% and is at the same time sensitive (concentrations in the range of 1 pmol/L miRNA can be reliably profiled). Because the novel approach did not require amplification steps, we obtained high reproducibility for up to 40 biological replicates. Importantly, our assay prototype exhibited a time to result of <3 h. With human blood samples, the assay was able to measure 4 miRNAs that can detect Alzheimer disease with a diagnostic accuracy of 82% and showed a Pearson correlation >0.994 with the gold standard qRT-PCR.
Our miRNA immunoassay allowed the measurement of miRNA signatures with sufficient analytical sensitivity and high specificity on commonly available laboratory equipment.</description><subject>Alzheimer Disease - blood</subject><subject>Alzheimer Disease - genetics</subject><subject>Alzheimer's disease</subject><subject>Antibodies, Monoclonal</subject><subject>Biomarkers</subject><subject>Biomarkers - blood</subject><subject>Blood banks</subject><subject>Deoxyribonucleic acid</subject><subject>Disease</subject><subject>DNA</subject><subject>Humans</subject><subject>Immunoassay</subject><subject>Immunoassay - instrumentation</subject><subject>Immunoassay - methods</subject><subject>Laboratories</subject><subject>Medical research</subject><subject>Methods</subject><subject>MicroRNAs - blood</subject><subject>Reproducibility of Results</subject><subject>Sensitivity and Specificity</subject><issn>0009-9147</issn><issn>1530-8561</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNqFkU1LAzEQhoMotlb_gciCFy9bM9l8HkvxC6qC6DnEbLam7EdNukL99aa09eDFwzAM87wvzLwInQMeQyGKa1v71n64Zkww0DEpCHB2gIbACpxLxuEQDTHGKldAxQCdxLhIIxWSH6MBSXtBCRsi9eht6F6eJplvsy-_Cl1WejNvu7jyNmZ99O08803Tt52J0awz05p6_e1CPEVHlamjO9v1EXq7vXmd3uez57uH6WSWWyrxKmeYSiuUwiAVtoAdE5Y5Z6Q1XEhaFSAMZa50hBMnoTRcpqoqC6VSsmLFCF1tfZeh--xdXOnGR-vq2rSu66MGCelEEIT-j3IuFMecQEIv_6CLrg_ptg0lCgpEMpUouqXSj2IMrtLL4BsT1hqw3qSg9ynoTQp6m0KSXezM-_fGlb-i_duLH3qQg4Q</recordid><startdate>201504</startdate><enddate>201504</enddate><creator>Kappel, Andreas</creator><creator>Backes, Christina</creator><creator>Huang, Yiwei</creator><creator>Zafari, Sachli</creator><creator>Leidinger, Petra</creator><creator>Meder, Benjamin</creator><creator>Schwarz, Herbert</creator><creator>Gumbrecht, Walter</creator><creator>Meese, Eckart</creator><creator>Staehler, Cord F</creator><creator>Keller, Andreas</creator><general>Oxford University Press</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>3V.</scope><scope>4U-</scope><scope>7QO</scope><scope>7RV</scope><scope>7TM</scope><scope>7U7</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>BKSAR</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>NAPCQ</scope><scope>P64</scope><scope>PCBAR</scope><scope>PDBOC</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>RC3</scope><scope>S0X</scope><scope>7X8</scope></search><sort><creationdate>201504</creationdate><title>MicroRNA in vitro diagnostics using immunoassay analyzers</title><author>Kappel, Andreas ; Backes, Christina ; Huang, Yiwei ; Zafari, Sachli ; Leidinger, Petra ; Meder, Benjamin ; Schwarz, Herbert ; Gumbrecht, Walter ; Meese, Eckart ; Staehler, Cord F ; Keller, Andreas</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c480t-5048c79901890c10e57c5eea8ca6784f317a45ede262e81da68da6ffc1d998f53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Alzheimer Disease - 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Academic</collection><jtitle>Clinical chemistry (Baltimore, Md.)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kappel, Andreas</au><au>Backes, Christina</au><au>Huang, Yiwei</au><au>Zafari, Sachli</au><au>Leidinger, Petra</au><au>Meder, Benjamin</au><au>Schwarz, Herbert</au><au>Gumbrecht, Walter</au><au>Meese, Eckart</au><au>Staehler, Cord F</au><au>Keller, Andreas</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>MicroRNA in vitro diagnostics using immunoassay analyzers</atitle><jtitle>Clinical chemistry (Baltimore, Md.)</jtitle><addtitle>Clin Chem</addtitle><date>2015-04</date><risdate>2015</risdate><volume>61</volume><issue>4</issue><spage>600</spage><epage>607</epage><pages>600-607</pages><issn>0009-9147</issn><eissn>1530-8561</eissn><abstract>The implementation of new biomarkers into clinical practice is one of the most important areas in medical research. Besides their clinical impact, novel in vitro diagnostic markers promise to have a substantial effect on healthcare costs. Although numerous publications report the discovery of biomarkers, only a fraction of those markers are routinely used. One key challenge is a measurement system that is compatible with clinical workflows.
We designed a new immunoassay for microRNA (miRNA) quantification. The assay combines streptavidin-linked microparticles, a biotinylated catcher oligonucleotide complementary to a single miRNA species, and finally, a monoclonal antibody to DNA/RNA heterohybrids labeled with acridinium ester. Importantly, our assay runs on standard immunoassay analyzers. After a technical validation of the assay, we evaluated the clinical performance on 4 Alzheimer disease miRNAs.
Our assay has an analytical specificity of 99.4% and is at the same time sensitive (concentrations in the range of 1 pmol/L miRNA can be reliably profiled). Because the novel approach did not require amplification steps, we obtained high reproducibility for up to 40 biological replicates. Importantly, our assay prototype exhibited a time to result of <3 h. With human blood samples, the assay was able to measure 4 miRNAs that can detect Alzheimer disease with a diagnostic accuracy of 82% and showed a Pearson correlation >0.994 with the gold standard qRT-PCR.
Our miRNA immunoassay allowed the measurement of miRNA signatures with sufficient analytical sensitivity and high specificity on commonly available laboratory equipment.</abstract><cop>England</cop><pub>Oxford University Press</pub><pmid>25617425</pmid><doi>10.1373/clinchem.2014.232165</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Clinical chemistry (Baltimore, Md.), 2015-04, Vol.61 (4), p.600-607 |
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| language | eng |
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| source | Oxford University Press Journals All Titles (1996-Current); MEDLINE |
| subjects | Alzheimer Disease - blood Alzheimer Disease - genetics Alzheimer's disease Antibodies, Monoclonal Biomarkers Biomarkers - blood Blood banks Deoxyribonucleic acid Disease DNA Humans Immunoassay Immunoassay - instrumentation Immunoassay - methods Laboratories Medical research Methods MicroRNAs - blood Reproducibility of Results Sensitivity and Specificity |
| title | MicroRNA in vitro diagnostics using immunoassay analyzers |
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