A First Tetraplex Assay for the Simultaneous Quantification of Total α-Synuclein, Tau, β-Amyloid42 and DJ-1 in Human Cerebrospinal Fluid
Identifieur interne : 000458 ( Main/Exploration ); précédent : 000457; suivant : 000459A First Tetraplex Assay for the Simultaneous Quantification of Total α-Synuclein, Tau, β-Amyloid42 and DJ-1 in Human Cerebrospinal Fluid
Auteurs : Niels Kruse [Allemagne] ; Michael G. Schlossmacher [Canada] ; Walter J. Schulz-Schaeffer [Allemagne] ; Eugeen Vanmechelen [Belgique] ; Hugo Vanderstichele [Belgique] ; Omar M. El-Agnaf [Qatar] ; Brit Mollenhauer [Allemagne]Source :
- PLoS ONE [ 1932-6203 ] ; 2016.
English descriptors
- KwdEn :
- Alzheimer Disease (cerebrospinal fluid), Alzheimer Disease (diagnosis), Amyloid beta-Peptides (cerebrospinal fluid), Biomarkers (cerebrospinal fluid), Enzyme-Linked Immunosorbent Assay (methods), Enzyme-Linked Immunosorbent Assay (statistics & numerical data), Humans, Limit of Detection, Neurodegenerative Diseases (cerebrospinal fluid), Neurodegenerative Diseases (diagnosis), Parkinson Disease (cerebrospinal fluid), Parkinson Disease (diagnosis), Peptide Fragments (cerebrospinal fluid), Protein Deglycase DJ-1 (cerebrospinal fluid), Reproducibility of Results, alpha-Synuclein (cerebrospinal fluid), tau Proteins (cerebrospinal fluid).
- MESH :
- chemical , cerebrospinal fluid : Amyloid beta-Peptides, Biomarkers, Peptide Fragments, Protein Deglycase DJ-1, alpha-Synuclein, tau Proteins.
- cerebrospinal fluid : Alzheimer Disease, Neurodegenerative Diseases, Parkinson Disease.
- diagnosis : Alzheimer Disease, Neurodegenerative Diseases, Parkinson Disease.
- methods : Enzyme-Linked Immunosorbent Assay.
- statistics & numerical data : Enzyme-Linked Immunosorbent Assay.
- Humans, Limit of Detection, Reproducibility of Results.
Abstract
The quantification of four distinct proteins (α-synuclein, β-amyloid1-42, DJ-1, and total tau) in cerebrospinal fluid (CSF) has been proposed as a laboratory-based platform for the diagnosis of Parkinson’s disease (PD) and Alzheimer’s disease (AD). While there is some clinical utility in measuring these markers individually, their usage in routine clinical testing remains challenging, in part due to substantial overlap of concentrations between healthy controls and diseased subjects. In contrast, measurement of different analytes in a single sample from individual patients in parallel appears to considerably improve the accuracy of AD or PD diagnosis. Here, we report the development and initial characterization of a first, electrochemiluminescence-based multiplex immunoassay for the simultaneous quantification of all four proteins (‘tetraplex’) in as little as 50 μl of CSF. In analytical performance experiments, we assessed its sensitivity, spike-recovery rate, parallelism and dilution linearity as well as the intra- and inter-assay variability. Using our in-house calibrators, we recorded a lower limit of detection for α-synuclein, β-amyloid42, DJ-1, and t-tau of 1.95, 1.24, 5.63, and 4.05 pg/ml, respectively. The corresponding, linear concentration range covered >3 orders of magnitude. In diluted CSF samples (up to 1:4), spike-recovery rates ranged from a low of 55% for β-amyloid42 to a high of 98% for DJ-1. Hillslopes ranged from 1.03 to 1.30, and inter-assay variability demonstrated very high reproducibility. Our newly established tetraplex assay represents a significant technical advance for fluid-based biomarker studies in neurodegenerative disorders allowing the simultaneous measurement of four pivotal makers in single CSF specimens. It provides exceptional sensitivity, accuracy and speed.
Url:
DOI: 10.1371/journal.pone.0153564
PubMed: 27116005
PubMed Central: 4846093
Affiliations:
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Schlossmacher</name><affiliation wicri:level="1"><nlm:aff id="aff002"><addr-line>Program in Neuroscience and Division of Neurology, The Ottawa Hospital, University of Ottawa Brain and Mind Research Institute, Ottawa, ON, Canada</addr-line></nlm:aff><country xml:lang="fr">Canada</country><wicri:regionArea>Program in Neuroscience and Division of Neurology, The Ottawa Hospital, University of Ottawa Brain and Mind Research Institute, Ottawa, ON</wicri:regionArea><wicri:noRegion>ON</wicri:noRegion></affiliation></author><author><name sortKey="Schulz Schaeffer, Walter J" sort="Schulz Schaeffer, Walter J" uniqKey="Schulz Schaeffer W" first="Walter J." last="Schulz-Schaeffer">Walter J. 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El-Agnaf</name><affiliation wicri:level="1"><nlm:aff id="aff004"><addr-line>Neurological Disorders Center, Qatar Biomedical Research Institute, and College of Science and Engineering, Hamad Bin Khalifa University (HBKU), Education City, Qatar Foundation, Doha, Qatar</addr-line></nlm:aff><country xml:lang="fr">Qatar</country><wicri:regionArea>Neurological Disorders Center, Qatar Biomedical Research Institute, and College of Science and Engineering, Hamad Bin Khalifa University (HBKU), Education City, Qatar Foundation, Doha</wicri:regionArea><wicri:noRegion>Doha</wicri:noRegion></affiliation></author><author><name sortKey="Mollenhauer, Brit" sort="Mollenhauer, Brit" uniqKey="Mollenhauer B" first="Brit" last="Mollenhauer">Brit Mollenhauer</name><affiliation wicri:level="3"><nlm:aff id="aff005"><addr-line>Paracelsus-Elena-Klinik, Kassel, Germany and University Medical Center, Göttingen, Germany</addr-line></nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea>Paracelsus-Elena-Klinik, Kassel, Germany and University Medical Center, Göttingen</wicri:regionArea><placeName><region type="land" nuts="2">Basse-Saxe</region><settlement type="city">Göttingen</settlement></placeName></affiliation></author></analytic><series><title level="j">PLoS ONE</title><idno type="eISSN">1932-6203</idno><imprint><date when="2016">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Alzheimer Disease (cerebrospinal fluid)</term><term>Alzheimer Disease (diagnosis)</term><term>Amyloid beta-Peptides (cerebrospinal fluid)</term><term>Biomarkers (cerebrospinal fluid)</term><term>Enzyme-Linked Immunosorbent Assay (methods)</term><term>Enzyme-Linked Immunosorbent Assay (statistics & numerical data)</term><term>Humans</term><term>Limit of Detection</term><term>Neurodegenerative Diseases (cerebrospinal fluid)</term><term>Neurodegenerative Diseases (diagnosis)</term><term>Parkinson Disease (cerebrospinal fluid)</term><term>Parkinson Disease (diagnosis)</term><term>Peptide Fragments (cerebrospinal fluid)</term><term>Protein Deglycase DJ-1 (cerebrospinal fluid)</term><term>Reproducibility of Results</term><term>alpha-Synuclein (cerebrospinal fluid)</term><term>tau Proteins (cerebrospinal fluid)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="cerebrospinal fluid" xml:lang="en"><term>Amyloid beta-Peptides</term><term>Biomarkers</term><term>Peptide Fragments</term><term>Protein Deglycase DJ-1</term><term>alpha-Synuclein</term><term>tau Proteins</term></keywords><keywords scheme="MESH" qualifier="cerebrospinal fluid" xml:lang="en"><term>Alzheimer Disease</term><term>Neurodegenerative Diseases</term><term>Parkinson Disease</term></keywords><keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Alzheimer Disease</term><term>Neurodegenerative Diseases</term><term>Parkinson Disease</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Enzyme-Linked Immunosorbent Assay</term></keywords><keywords scheme="MESH" qualifier="statistics & numerical data" xml:lang="en"><term>Enzyme-Linked Immunosorbent Assay</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Humans</term><term>Limit of Detection</term><term>Reproducibility of Results</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>The quantification of four distinct proteins (α-synuclein, β-amyloid<sub>1-42</sub>, DJ-1, and total tau) in cerebrospinal fluid (CSF) has been proposed as a laboratory-based platform for the diagnosis of Parkinson’s disease (PD) and Alzheimer’s disease (AD). While there is some clinical utility in measuring these markers individually, their usage in routine clinical testing remains challenging, in part due to substantial overlap of concentrations between healthy controls and diseased subjects. In contrast, measurement of different analytes in a single sample from individual patients in parallel appears to considerably improve the accuracy of AD or PD diagnosis. Here, we report the development and initial characterization of a first, electrochemiluminescence-based multiplex immunoassay for the simultaneous quantification of all four proteins (‘tetraplex’) in as little as 50 μl of CSF. In analytical performance experiments, we assessed its sensitivity, spike-recovery rate, parallelism and dilution linearity as well as the intra- and inter-assay variability. Using our in-house calibrators, we recorded a lower limit of detection for α-synuclein, β-amyloid<sub>42</sub>, DJ-1, and t-tau of 1.95, 1.24, 5.63, and 4.05 pg/ml, respectively. The corresponding, linear concentration range covered >3 orders of magnitude. In diluted CSF samples (up to 1:4), spike-recovery rates ranged from a low of 55% for β-amyloid<sub>42</sub> to a high of 98% for DJ-1. Hillslopes ranged from 1.03 to 1.30, and inter-assay variability demonstrated very high reproducibility. Our newly established tetraplex assay represents a significant technical advance for fluid-based biomarker studies in neurodegenerative disorders allowing the simultaneous measurement of four pivotal makers in single CSF specimens. It provides exceptional sensitivity, accuracy and speed.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Shi, M" uniqKey="Shi M">M Shi</name></author><author><name sortKey="Bradner, J" uniqKey="Bradner J">J Bradner</name></author><author><name sortKey="Hancock, Am" uniqKey="Hancock A">AM Hancock</name></author><author><name sortKey="Chung, Ka" uniqKey="Chung K">KA Chung</name></author><author><name sortKey="Quinn, Jf" uniqKey="Quinn J">JF Quinn</name></author><author><name sortKey="Peskind, Er" uniqKey="Peskind E">ER Peskind</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Abdi, F" uniqKey="Abdi F">F Abdi</name></author><author><name sortKey="Quinn, Jf" uniqKey="Quinn J">JF Quinn</name></author><author><name sortKey="Jankovic, J" uniqKey="Jankovic J">J Jankovic</name></author><author><name sortKey="Mcintosh, M" uniqKey="Mcintosh M">M McIntosh</name></author><author><name sortKey="Leverenz, Jb" 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Nielsen</name></author><author><name sortKey="Hojrup, P" uniqKey="Hojrup P">P Hojrup</name></author><author><name sortKey="Gliemann, J" uniqKey="Gliemann J">J Gliemann</name></author><author><name sortKey="Jakes, R" uniqKey="Jakes R">R Jakes</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Clinton, Lk" uniqKey="Clinton L">LK Clinton</name></author><author><name sortKey="Blurton Jones, M" uniqKey="Blurton Jones M">M Blurton-Jones</name></author><author><name sortKey="Myczek, K" uniqKey="Myczek K">K Myczek</name></author><author><name sortKey="Trojanowski, Jq" uniqKey="Trojanowski J">JQ Trojanowski</name></author><author><name sortKey="Laferla, Fm" uniqKey="Laferla F">FM LaFerla</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Mattsson, N" uniqKey="Mattsson N">N Mattsson</name></author><author><name sortKey="Andreasson, U" uniqKey="Andreasson U">U Andreasson</name></author><author><name sortKey="Persson, S" uniqKey="Persson S">S Persson</name></author><author><name sortKey="Arai, H" uniqKey="Arai H">H Arai</name></author><author><name sortKey="Batish, Sd" uniqKey="Batish S">SD Batish</name></author><author><name sortKey="Bernardini, S" uniqKey="Bernardini S">S Bernardini</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Kruse, N" uniqKey="Kruse N">N Kruse</name></author><author><name sortKey="Persson, S" uniqKey="Persson S">S Persson</name></author><author><name sortKey="Bahl, Jmc" uniqKey="Bahl J">JMC Bahl</name></author><author><name sortKey="Baldeiras, I" uniqKey="Baldeiras I">I Baldeiras</name></author><author><name sortKey="Capello, E" uniqKey="Capello E">E Capello</name></author><author><name sortKey="Bocchio Chiavetto, L" uniqKey="Bocchio Chiavetto L">L Bocchio Chiavetto</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Montine, Tj" uniqKey="Montine T">TJ Montine</name></author><author><name sortKey="Shi, M" uniqKey="Shi M">M Shi</name></author><author><name sortKey="Quinn, Jf" uniqKey="Quinn J">JF Quinn</name></author><author><name sortKey="Peskind, Er" uniqKey="Peskind E">ER Peskind</name></author><author><name sortKey="Craft, S" uniqKey="Craft S">S Craft</name></author><author><name sortKey="Ginghina, C" uniqKey="Ginghina C">C Ginghina</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list><country><li>Allemagne</li><li>Belgique</li><li>Canada</li><li>Qatar</li></country><region><li>Basse-Saxe</li></region><settlement><li>Göttingen</li></settlement></list><tree><country name="Allemagne"><region name="Basse-Saxe"><name sortKey="Kruse, Niels" sort="Kruse, Niels" uniqKey="Kruse N" first="Niels" last="Kruse">Niels Kruse</name></region><name sortKey="Mollenhauer, Brit" sort="Mollenhauer, Brit" uniqKey="Mollenhauer B" first="Brit" last="Mollenhauer">Brit Mollenhauer</name><name sortKey="Schulz Schaeffer, Walter J" sort="Schulz Schaeffer, Walter J" uniqKey="Schulz Schaeffer W" first="Walter J." last="Schulz-Schaeffer">Walter J. Schulz-Schaeffer</name></country><country name="Canada"><noRegion><name sortKey="Schlossmacher, Michael G" sort="Schlossmacher, Michael G" uniqKey="Schlossmacher M" first="Michael G." last="Schlossmacher">Michael G. Schlossmacher</name></noRegion></country><country name="Belgique"><noRegion><name sortKey="Vanmechelen, Eugeen" sort="Vanmechelen, Eugeen" uniqKey="Vanmechelen E" first="Eugeen" last="Vanmechelen">Eugeen Vanmechelen</name></noRegion><name sortKey="Vanderstichele, Hugo" sort="Vanderstichele, Hugo" uniqKey="Vanderstichele H" first="Hugo" last="Vanderstichele">Hugo Vanderstichele</name></country><country name="Qatar"><noRegion><name sortKey="El Agnaf, Omar M" sort="El Agnaf, Omar M" uniqKey="El Agnaf O" first="Omar M." last="El-Agnaf">Omar M. El-Agnaf</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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