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Article: Quantitative analysis of circulating mitochondrial DNA in plasma

TitleQuantitative analysis of circulating mitochondrial DNA in plasma
Authors
Issue Date2003
PublisherAmerican Association for Clinical Chemistry, Inc. The Journal's web site is located at http://www.clinchem.org
Citation
Clinical Chemistry, 2003, v. 49 n. 5, p. 719-726 How to Cite?
AbstractBackground: Recent studies have demonstrated the existence of circulating mitochondrial DNA in plasma and serum, but the concentrations and physical characteristics of circulating mitochondrial DNA are unknown. The aim of this study was to develop an assay to quantify mitochondrial DNA in the plasma of healthy individuals. Methods: We adopted a real-time quantitative PCR approach and evaluated the specificity of the assay for detecting mitochondrial DNA with a cell line (ρ 0) devoid of mitochondria. The concentrations and physical characteristics of circulating mitochondrial DNA were investigated by experiments conducted in three modules. In module 1, we evaluated the concentrations of mitochondrial DNA in plasma aliquots derived from four blood-processing protocols. In module 2, we investigated the existence of both particle-associated and free forms of mitochondrial DNA in plasma by subjecting plasma to filtration and ultracentrifugation. In module 3, we used filters with different pore sizes to investigate the size characteristics of the particle-associated fraction of circulating mitochondrial DNA. Results: The mitochondrial DNA-specific, real-time quantitative PCR had a dynamic range of five orders of magnitude and a sensitivity that enabled detection of one copy of mitochondrial DNA in plasma. In module 1, we found significant differences in the amounts of circulating mitochondrial DNA among plasma aliquots processed by different methods. Data from module 2 revealed that a significant fraction of mitochondrial DNA in plasma was filterable or pelletable by ultracentrifugation. Module 3 demonstrated that filters with different pore sizes removed mitochondrial DNA from plasma to different degrees. Conclusions: Both particle-associated and free mitochondrial DNA are present in plasma, and their respective concentrations are affected by the process used to harvest plasma from whole blood. These results may have implications in the design of future studies on circulating mitochondrial DNA measured in different disease conditions. © 2003 American Association for Clinical Chemistry.
Persistent Identifierhttp://hdl.handle.net/10722/172824
ISSN
2015 Impact Factor: 7.457
2015 SCImago Journal Rankings: 2.472
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorChiu, RWKen_US
dc.contributor.authorChan, LYSen_US
dc.contributor.authorLam, NYLen_US
dc.contributor.authorTsui, NBYen_US
dc.contributor.authorNg, EKOen_US
dc.contributor.authorRainer, THen_US
dc.contributor.authorLo, YMDen_US
dc.date.accessioned2012-10-30T06:25:07Z-
dc.date.available2012-10-30T06:25:07Z-
dc.date.issued2003en_US
dc.identifier.citationClinical Chemistry, 2003, v. 49 n. 5, p. 719-726en_US
dc.identifier.issn0009-9147en_US
dc.identifier.urihttp://hdl.handle.net/10722/172824-
dc.description.abstractBackground: Recent studies have demonstrated the existence of circulating mitochondrial DNA in plasma and serum, but the concentrations and physical characteristics of circulating mitochondrial DNA are unknown. The aim of this study was to develop an assay to quantify mitochondrial DNA in the plasma of healthy individuals. Methods: We adopted a real-time quantitative PCR approach and evaluated the specificity of the assay for detecting mitochondrial DNA with a cell line (ρ 0) devoid of mitochondria. The concentrations and physical characteristics of circulating mitochondrial DNA were investigated by experiments conducted in three modules. In module 1, we evaluated the concentrations of mitochondrial DNA in plasma aliquots derived from four blood-processing protocols. In module 2, we investigated the existence of both particle-associated and free forms of mitochondrial DNA in plasma by subjecting plasma to filtration and ultracentrifugation. In module 3, we used filters with different pore sizes to investigate the size characteristics of the particle-associated fraction of circulating mitochondrial DNA. Results: The mitochondrial DNA-specific, real-time quantitative PCR had a dynamic range of five orders of magnitude and a sensitivity that enabled detection of one copy of mitochondrial DNA in plasma. In module 1, we found significant differences in the amounts of circulating mitochondrial DNA among plasma aliquots processed by different methods. Data from module 2 revealed that a significant fraction of mitochondrial DNA in plasma was filterable or pelletable by ultracentrifugation. Module 3 demonstrated that filters with different pore sizes removed mitochondrial DNA from plasma to different degrees. Conclusions: Both particle-associated and free mitochondrial DNA are present in plasma, and their respective concentrations are affected by the process used to harvest plasma from whole blood. These results may have implications in the design of future studies on circulating mitochondrial DNA measured in different disease conditions. © 2003 American Association for Clinical Chemistry.en_US
dc.languageengen_US
dc.publisherAmerican Association for Clinical Chemistry, Inc. The Journal's web site is located at http://www.clinchem.orgen_US
dc.relation.ispartofClinical Chemistryen_US
dc.subject.meshBlood Circulationen_US
dc.subject.meshBlood Specimen Collectionen_US
dc.subject.meshDna, Mitochondrial - Blooden_US
dc.subject.meshHumansen_US
dc.subject.meshLinear Modelsen_US
dc.subject.meshPolymerase Chain Reactionen_US
dc.subject.meshSensitivity And Specificityen_US
dc.titleQuantitative analysis of circulating mitochondrial DNA in plasmaen_US
dc.typeArticleen_US
dc.identifier.emailNg, EKO: ngko@hku.hken_US
dc.identifier.authorityNg, EKO=rp01364en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1373/49.5.719en_US
dc.identifier.pmid12709361-
dc.identifier.scopuseid_2-s2.0-0037407143en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0037407143&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume49en_US
dc.identifier.issue5en_US
dc.identifier.spage719en_US
dc.identifier.epage726en_US
dc.identifier.isiWOS:000182541900002-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridChiu, RWK=7103038413en_US
dc.identifier.scopusauthoridChan, LYS=8108378300en_US
dc.identifier.scopusauthoridLam, NYL=7101750743en_US
dc.identifier.scopusauthoridTsui, NBY=6602401748en_US
dc.identifier.scopusauthoridNg, EKO=21135553700en_US
dc.identifier.scopusauthoridRainer, TH=7004489495en_US
dc.identifier.scopusauthoridLo, YMD=7401935391en_US

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