Using a quantitative quadruple immunofluorescent assay to diagnose isolated mitochondrial Complex I deficiency
Isolated Complex I (CI) deficiency is the most commonly observed mitochondrial respiratory chain biochemical defect, affecting the largest OXPHOS component. CI is genetically heterogeneous; pathogenic variants affect one of 38 nuclear-encoded subunits, 7 mitochondrial DNA (mtDNA)-encoded subunits or...
Gespeichert in:
| Veröffentlicht in: | Scientific reports 2017-11, Vol.7 (1), p.15676-13, Article 15676 |
|---|---|
| Hauptverfasser: | , , , , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 13 |
|---|---|
| container_issue | 1 |
| container_start_page | 15676 |
| container_title | Scientific reports |
| container_volume | 7 |
| creator | Ahmed, Syeda T. Alston, Charlotte L. Hopton, Sila He, Langping Hargreaves, Iain P. Falkous, Gavin Oláhová, Monika McFarland, Robert Turnbull, Doug M. Rocha, Mariana C. Taylor, Robert W. |
| description | Isolated Complex I (CI) deficiency is the most commonly observed mitochondrial respiratory chain biochemical defect, affecting the largest OXPHOS component. CI is genetically heterogeneous; pathogenic variants affect one of 38 nuclear-encoded subunits, 7 mitochondrial DNA (mtDNA)-encoded subunits or 14 known CI assembly factors. The laboratory diagnosis relies on the spectrophotometric assay of enzyme activity in mitochondrially-enriched tissue homogenates, requiring at least 50 mg skeletal muscle, as there is no reliable histochemical method for assessing CI activity directly in tissue cryosections. We have assessed a validated quadruple immunofluorescent OXPHOS (IHC) assay to detect CI deficiency in the diagnostic setting, using 10 µm transverse muscle sections from 25 patients with genetically-proven pathogenic CI variants. We observed loss of NDUFB8 immunoreactivity in all patients with mutations affecting nuclear-encoding structural subunits and assembly factors, whilst only 3 of the 10 patients with mutations affecting mtDNA-encoded structural subunits showed loss of NDUFB8, confirmed by BN-PAGE analysis of CI assembly and IHC using an alternative, commercially-available CI (NDUFS3) antibody. The IHC assay has clear diagnostic potential to identify patients with a CI defect of Mendelian origins, whilst highlighting the necessity of complete mitochondrial genome sequencing in the diagnostic work-up of patients with suspected mitochondrial disease. |
| doi_str_mv | 10.1038/s41598-017-14623-2 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_5688115</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>1964559624</sourcerecordid><originalsourceid>FETCH-LOGICAL-c474t-3ff4d93c654373fd54ad90d90e4654844c0670c07c097122db1c33099a53574c3</originalsourceid><addsrcrecordid>eNp1kVFPHCEQx0lTo0b9Aj40JL70ZRUG2F1eTMylrSYmfanPBIE9MbtwAmt6376cp-Y0KSFhYH7zZyZ_hE4pOaeE9ReZUyH7htCuobwF1sAXdAiEiwYYwNed-ACd5PxI6hIgOZX76AAk5QCiO0ThLvuwxBo_zToUX3Txz25zsWlejQ77aZpDHMY5JpeNCwXrnPUal4it18sQc2VyHHVxFk--RPMQg01ej3gRp6rwF99g6wZvvAtmfYz2Bj1md_J6HqG7nz_-LK6b29-_bhZXt43hHS8NGwZuJTOt4KxjgxVcW0nqdrw-9Zwb0nbEkM4Q2VEAe08NY0RKLZjouGFH6HKru5rvJ2c3jSc9qlXyk05rFbVXHzPBP6hlfFai7XtKRRX4_iqQ4tPsclGTr_OPow4uzllR2QrgAgit6Nkn9DHOKdTxNhQXQrbAKwVbyqSYc3LDezOUqI2jauuoqo6qF0cV1KJvu2O8l7z5VwG2BXJNhaVLO3__X_YfhrStiw</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1964559624</pqid></control><display><type>article</type><title>Using a quantitative quadruple immunofluorescent assay to diagnose isolated mitochondrial Complex I deficiency</title><source>MEDLINE</source><source>DOAJ Directory of Open Access Journals</source><source>Springer Nature OA Free Journals</source><source>Nature Free</source><source>EZB-FREE-00999 freely available EZB journals</source><source>PubMed Central</source><source>Free Full-Text Journals in Chemistry</source><creator>Ahmed, Syeda T. ; Alston, Charlotte L. ; Hopton, Sila ; He, Langping ; Hargreaves, Iain P. ; Falkous, Gavin ; Oláhová, Monika ; McFarland, Robert ; Turnbull, Doug M. ; Rocha, Mariana C. ; Taylor, Robert W.</creator><creatorcontrib>Ahmed, Syeda T. ; Alston, Charlotte L. ; Hopton, Sila ; He, Langping ; Hargreaves, Iain P. ; Falkous, Gavin ; Oláhová, Monika ; McFarland, Robert ; Turnbull, Doug M. ; Rocha, Mariana C. ; Taylor, Robert W.</creatorcontrib><description>Isolated Complex I (CI) deficiency is the most commonly observed mitochondrial respiratory chain biochemical defect, affecting the largest OXPHOS component. CI is genetically heterogeneous; pathogenic variants affect one of 38 nuclear-encoded subunits, 7 mitochondrial DNA (mtDNA)-encoded subunits or 14 known CI assembly factors. The laboratory diagnosis relies on the spectrophotometric assay of enzyme activity in mitochondrially-enriched tissue homogenates, requiring at least 50 mg skeletal muscle, as there is no reliable histochemical method for assessing CI activity directly in tissue cryosections. We have assessed a validated quadruple immunofluorescent OXPHOS (IHC) assay to detect CI deficiency in the diagnostic setting, using 10 µm transverse muscle sections from 25 patients with genetically-proven pathogenic CI variants. We observed loss of NDUFB8 immunoreactivity in all patients with mutations affecting nuclear-encoding structural subunits and assembly factors, whilst only 3 of the 10 patients with mutations affecting mtDNA-encoded structural subunits showed loss of NDUFB8, confirmed by BN-PAGE analysis of CI assembly and IHC using an alternative, commercially-available CI (NDUFS3) antibody. The IHC assay has clear diagnostic potential to identify patients with a CI defect of Mendelian origins, whilst highlighting the necessity of complete mitochondrial genome sequencing in the diagnostic work-up of patients with suspected mitochondrial disease.</description><identifier>ISSN: 2045-2322</identifier><identifier>EISSN: 2045-2322</identifier><identifier>DOI: 10.1038/s41598-017-14623-2</identifier><identifier>PMID: 29142257</identifier><language>eng</language><publisher>London: Nature Publishing Group UK</publisher><subject>13/1 ; 13/51 ; 45 ; 45/23 ; 631/208/1516 ; 631/45/881 ; 82 ; 82/29 ; Biopsy ; Cell Nucleus - genetics ; Child ; Child, Preschool ; DNA, Mitochondrial - genetics ; Electron transport ; Electron transport chain ; Electron Transport Complex I - deficiency ; Electron Transport Complex I - genetics ; Enzymatic activity ; Female ; Fluorescent Antibody Technique ; Fluoroimmunoassay - methods ; Genetic Heterogeneity ; Genomes ; Humanities and Social Sciences ; Humans ; Immunoreactivity ; Male ; Mitochondria - genetics ; Mitochondrial Diseases - diagnosis ; Mitochondrial Diseases - genetics ; Mitochondrial Diseases - pathology ; Mitochondrial DNA ; multidisciplinary ; Muscle, Skeletal - metabolism ; Muscle, Skeletal - pathology ; Mutation ; NADH Dehydrogenase - genetics ; NADH-ubiquinone oxidoreductase ; Oxidative Phosphorylation ; Science ; Science (multidisciplinary) ; Skeletal muscle ; Spectrophotometry</subject><ispartof>Scientific reports, 2017-11, Vol.7 (1), p.15676-13, Article 15676</ispartof><rights>The Author(s) 2017</rights><rights>2017. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c474t-3ff4d93c654373fd54ad90d90e4654844c0670c07c097122db1c33099a53574c3</citedby><cites>FETCH-LOGICAL-c474t-3ff4d93c654373fd54ad90d90e4654844c0670c07c097122db1c33099a53574c3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5688115/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5688115/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,27924,27925,41120,42189,51576,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29142257$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ahmed, Syeda T.</creatorcontrib><creatorcontrib>Alston, Charlotte L.</creatorcontrib><creatorcontrib>Hopton, Sila</creatorcontrib><creatorcontrib>He, Langping</creatorcontrib><creatorcontrib>Hargreaves, Iain P.</creatorcontrib><creatorcontrib>Falkous, Gavin</creatorcontrib><creatorcontrib>Oláhová, Monika</creatorcontrib><creatorcontrib>McFarland, Robert</creatorcontrib><creatorcontrib>Turnbull, Doug M.</creatorcontrib><creatorcontrib>Rocha, Mariana C.</creatorcontrib><creatorcontrib>Taylor, Robert W.</creatorcontrib><title>Using a quantitative quadruple immunofluorescent assay to diagnose isolated mitochondrial Complex I deficiency</title><title>Scientific reports</title><addtitle>Sci Rep</addtitle><addtitle>Sci Rep</addtitle><description>Isolated Complex I (CI) deficiency is the most commonly observed mitochondrial respiratory chain biochemical defect, affecting the largest OXPHOS component. CI is genetically heterogeneous; pathogenic variants affect one of 38 nuclear-encoded subunits, 7 mitochondrial DNA (mtDNA)-encoded subunits or 14 known CI assembly factors. The laboratory diagnosis relies on the spectrophotometric assay of enzyme activity in mitochondrially-enriched tissue homogenates, requiring at least 50 mg skeletal muscle, as there is no reliable histochemical method for assessing CI activity directly in tissue cryosections. We have assessed a validated quadruple immunofluorescent OXPHOS (IHC) assay to detect CI deficiency in the diagnostic setting, using 10 µm transverse muscle sections from 25 patients with genetically-proven pathogenic CI variants. We observed loss of NDUFB8 immunoreactivity in all patients with mutations affecting nuclear-encoding structural subunits and assembly factors, whilst only 3 of the 10 patients with mutations affecting mtDNA-encoded structural subunits showed loss of NDUFB8, confirmed by BN-PAGE analysis of CI assembly and IHC using an alternative, commercially-available CI (NDUFS3) antibody. The IHC assay has clear diagnostic potential to identify patients with a CI defect of Mendelian origins, whilst highlighting the necessity of complete mitochondrial genome sequencing in the diagnostic work-up of patients with suspected mitochondrial disease.</description><subject>13/1</subject><subject>13/51</subject><subject>45</subject><subject>45/23</subject><subject>631/208/1516</subject><subject>631/45/881</subject><subject>82</subject><subject>82/29</subject><subject>Biopsy</subject><subject>Cell Nucleus - genetics</subject><subject>Child</subject><subject>Child, Preschool</subject><subject>DNA, Mitochondrial - genetics</subject><subject>Electron transport</subject><subject>Electron transport chain</subject><subject>Electron Transport Complex I - deficiency</subject><subject>Electron Transport Complex I - genetics</subject><subject>Enzymatic activity</subject><subject>Female</subject><subject>Fluorescent Antibody Technique</subject><subject>Fluoroimmunoassay - methods</subject><subject>Genetic Heterogeneity</subject><subject>Genomes</subject><subject>Humanities and Social Sciences</subject><subject>Humans</subject><subject>Immunoreactivity</subject><subject>Male</subject><subject>Mitochondria - genetics</subject><subject>Mitochondrial Diseases - diagnosis</subject><subject>Mitochondrial Diseases - genetics</subject><subject>Mitochondrial Diseases - pathology</subject><subject>Mitochondrial DNA</subject><subject>multidisciplinary</subject><subject>Muscle, Skeletal - metabolism</subject><subject>Muscle, Skeletal - pathology</subject><subject>Mutation</subject><subject>NADH Dehydrogenase - genetics</subject><subject>NADH-ubiquinone oxidoreductase</subject><subject>Oxidative Phosphorylation</subject><subject>Science</subject><subject>Science (multidisciplinary)</subject><subject>Skeletal muscle</subject><subject>Spectrophotometry</subject><issn>2045-2322</issn><issn>2045-2322</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>C6C</sourceid><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp1kVFPHCEQx0lTo0b9Aj40JL70ZRUG2F1eTMylrSYmfanPBIE9MbtwAmt6376cp-Y0KSFhYH7zZyZ_hE4pOaeE9ReZUyH7htCuobwF1sAXdAiEiwYYwNed-ACd5PxI6hIgOZX76AAk5QCiO0ThLvuwxBo_zToUX3Txz25zsWlejQ77aZpDHMY5JpeNCwXrnPUal4it18sQc2VyHHVxFk--RPMQg01ej3gRp6rwF99g6wZvvAtmfYz2Bj1md_J6HqG7nz_-LK6b29-_bhZXt43hHS8NGwZuJTOt4KxjgxVcW0nqdrw-9Zwb0nbEkM4Q2VEAe08NY0RKLZjouGFH6HKru5rvJ2c3jSc9qlXyk05rFbVXHzPBP6hlfFai7XtKRRX4_iqQ4tPsclGTr_OPow4uzllR2QrgAgit6Nkn9DHOKdTxNhQXQrbAKwVbyqSYc3LDezOUqI2jauuoqo6qF0cV1KJvu2O8l7z5VwG2BXJNhaVLO3__X_YfhrStiw</recordid><startdate>20171115</startdate><enddate>20171115</enddate><creator>Ahmed, Syeda T.</creator><creator>Alston, Charlotte L.</creator><creator>Hopton, Sila</creator><creator>He, Langping</creator><creator>Hargreaves, Iain P.</creator><creator>Falkous, Gavin</creator><creator>Oláhová, Monika</creator><creator>McFarland, Robert</creator><creator>Turnbull, Doug M.</creator><creator>Rocha, Mariana C.</creator><creator>Taylor, Robert W.</creator><general>Nature Publishing Group UK</general><general>Nature Publishing Group</general><scope>C6C</scope><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>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7P</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20171115</creationdate><title>Using a quantitative quadruple immunofluorescent assay to diagnose isolated mitochondrial Complex I deficiency</title><author>Ahmed, Syeda T. ; Alston, Charlotte L. ; Hopton, Sila ; He, Langping ; Hargreaves, Iain P. ; Falkous, Gavin ; Oláhová, Monika ; McFarland, Robert ; Turnbull, Doug M. ; Rocha, Mariana C. ; Taylor, Robert W.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c474t-3ff4d93c654373fd54ad90d90e4654844c0670c07c097122db1c33099a53574c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>13/1</topic><topic>13/51</topic><topic>45</topic><topic>45/23</topic><topic>631/208/1516</topic><topic>631/45/881</topic><topic>82</topic><topic>82/29</topic><topic>Biopsy</topic><topic>Cell Nucleus - genetics</topic><topic>Child</topic><topic>Child, Preschool</topic><topic>DNA, Mitochondrial - genetics</topic><topic>Electron transport</topic><topic>Electron transport chain</topic><topic>Electron Transport Complex I - deficiency</topic><topic>Electron Transport Complex I - genetics</topic><topic>Enzymatic activity</topic><topic>Female</topic><topic>Fluorescent Antibody Technique</topic><topic>Fluoroimmunoassay - methods</topic><topic>Genetic Heterogeneity</topic><topic>Genomes</topic><topic>Humanities and Social Sciences</topic><topic>Humans</topic><topic>Immunoreactivity</topic><topic>Male</topic><topic>Mitochondria - genetics</topic><topic>Mitochondrial Diseases - diagnosis</topic><topic>Mitochondrial Diseases - genetics</topic><topic>Mitochondrial Diseases - pathology</topic><topic>Mitochondrial DNA</topic><topic>multidisciplinary</topic><topic>Muscle, Skeletal - metabolism</topic><topic>Muscle, Skeletal - pathology</topic><topic>Mutation</topic><topic>NADH Dehydrogenase - genetics</topic><topic>NADH-ubiquinone oxidoreductase</topic><topic>Oxidative Phosphorylation</topic><topic>Science</topic><topic>Science (multidisciplinary)</topic><topic>Skeletal muscle</topic><topic>Spectrophotometry</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ahmed, Syeda T.</creatorcontrib><creatorcontrib>Alston, Charlotte L.</creatorcontrib><creatorcontrib>Hopton, Sila</creatorcontrib><creatorcontrib>He, Langping</creatorcontrib><creatorcontrib>Hargreaves, Iain P.</creatorcontrib><creatorcontrib>Falkous, Gavin</creatorcontrib><creatorcontrib>Oláhová, Monika</creatorcontrib><creatorcontrib>McFarland, Robert</creatorcontrib><creatorcontrib>Turnbull, Doug M.</creatorcontrib><creatorcontrib>Rocha, Mariana C.</creatorcontrib><creatorcontrib>Taylor, Robert W.</creatorcontrib><collection>Springer Nature OA Free Journals</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database</collection><collection>Biological Science Database</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Scientific reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ahmed, Syeda T.</au><au>Alston, Charlotte L.</au><au>Hopton, Sila</au><au>He, Langping</au><au>Hargreaves, Iain P.</au><au>Falkous, Gavin</au><au>Oláhová, Monika</au><au>McFarland, Robert</au><au>Turnbull, Doug M.</au><au>Rocha, Mariana C.</au><au>Taylor, Robert W.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Using a quantitative quadruple immunofluorescent assay to diagnose isolated mitochondrial Complex I deficiency</atitle><jtitle>Scientific reports</jtitle><stitle>Sci Rep</stitle><addtitle>Sci Rep</addtitle><date>2017-11-15</date><risdate>2017</risdate><volume>7</volume><issue>1</issue><spage>15676</spage><epage>13</epage><pages>15676-13</pages><artnum>15676</artnum><issn>2045-2322</issn><eissn>2045-2322</eissn><abstract>Isolated Complex I (CI) deficiency is the most commonly observed mitochondrial respiratory chain biochemical defect, affecting the largest OXPHOS component. CI is genetically heterogeneous; pathogenic variants affect one of 38 nuclear-encoded subunits, 7 mitochondrial DNA (mtDNA)-encoded subunits or 14 known CI assembly factors. The laboratory diagnosis relies on the spectrophotometric assay of enzyme activity in mitochondrially-enriched tissue homogenates, requiring at least 50 mg skeletal muscle, as there is no reliable histochemical method for assessing CI activity directly in tissue cryosections. We have assessed a validated quadruple immunofluorescent OXPHOS (IHC) assay to detect CI deficiency in the diagnostic setting, using 10 µm transverse muscle sections from 25 patients with genetically-proven pathogenic CI variants. We observed loss of NDUFB8 immunoreactivity in all patients with mutations affecting nuclear-encoding structural subunits and assembly factors, whilst only 3 of the 10 patients with mutations affecting mtDNA-encoded structural subunits showed loss of NDUFB8, confirmed by BN-PAGE analysis of CI assembly and IHC using an alternative, commercially-available CI (NDUFS3) antibody. The IHC assay has clear diagnostic potential to identify patients with a CI defect of Mendelian origins, whilst highlighting the necessity of complete mitochondrial genome sequencing in the diagnostic work-up of patients with suspected mitochondrial disease.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>29142257</pmid><doi>10.1038/s41598-017-14623-2</doi><tpages>13</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 2045-2322 |
| ispartof | Scientific reports, 2017-11, Vol.7 (1), p.15676-13, Article 15676 |
| issn | 2045-2322 2045-2322 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_5688115 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Springer Nature OA Free Journals; Nature Free; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | 13/1 13/51 45 45/23 631/208/1516 631/45/881 82 82/29 Biopsy Cell Nucleus - genetics Child Child, Preschool DNA, Mitochondrial - genetics Electron transport Electron transport chain Electron Transport Complex I - deficiency Electron Transport Complex I - genetics Enzymatic activity Female Fluorescent Antibody Technique Fluoroimmunoassay - methods Genetic Heterogeneity Genomes Humanities and Social Sciences Humans Immunoreactivity Male Mitochondria - genetics Mitochondrial Diseases - diagnosis Mitochondrial Diseases - genetics Mitochondrial Diseases - pathology Mitochondrial DNA multidisciplinary Muscle, Skeletal - metabolism Muscle, Skeletal - pathology Mutation NADH Dehydrogenase - genetics NADH-ubiquinone oxidoreductase Oxidative Phosphorylation Science Science (multidisciplinary) Skeletal muscle Spectrophotometry |
| title | Using a quantitative quadruple immunofluorescent assay to diagnose isolated mitochondrial Complex I deficiency |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-27T22%3A03%3A14IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Using%20a%20quantitative%20quadruple%20immunofluorescent%20assay%20to%20diagnose%20isolated%20mitochondrial%20Complex%20I%20deficiency&rft.jtitle=Scientific%20reports&rft.au=Ahmed,%20Syeda%20T.&rft.date=2017-11-15&rft.volume=7&rft.issue=1&rft.spage=15676&rft.epage=13&rft.pages=15676-13&rft.artnum=15676&rft.issn=2045-2322&rft.eissn=2045-2322&rft_id=info:doi/10.1038/s41598-017-14623-2&rft_dat=%3Cproquest_pubme%3E1964559624%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1964559624&rft_id=info:pmid/29142257&rfr_iscdi=true |