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Article: Stability of endogenous and added RNA in blood specimens, serum, and plasma

TitleStability of endogenous and added RNA in blood specimens, serum, and plasma
Authors
Issue Date2002
PublisherAmerican Association for Clinical Chemistry, Inc. The Journal's web site is located at http://www.clinchem.org
Citation
Clinical Chemistry, 2002, v. 48 n. 10, p. 1647-1653 How to Cite?
AbstractBackground: Circulating RNA in plasma/serum is an emerging field for noninvasive molecular diagnosis. Because RNA is widely thought to be labile in the circulation, we investigated the stability and various preanalytical factors that may affect RNA concentrations in blood specimens. Methods: Blood samples were collected from 65 healthy volunteers. The effects of two preanalytical variables Were studied: (a) time delay in processing of EDTA blood and clotted blood after venesection, and (b) freezing and thawing of plasma and serum. The lability of free added RNA in plasma was also investigated. Plasma/serum RNA was measured by a real-time quantitative reverse transcription-PCR assay for glyceraldehyde 3-phosphate dehydrogenase mRNA, whereas DNA was measured by a real-time quantitative PCR assay for the β-globin gene. Results: No significant difference was found for plasma RNA concentrations obtained from uncentrifuged EDTA blood that had been left at 4 °C for 0, 6, and 24 h (P = 0.182). On the other hand, the serum RNA concentrations increased significantly over 24 h when uncentrifuged clotted blood was stored at 4 °C (P <0.05). In comparison, >99% of the free added RNA could no longer be amplified after incubation in plasma for 15 s. Never-frozen plasma, freeze-thawed plasma, and thawed plasma left at room temperature for 1 h showed no significant differences in RNA concentration (P = 0:465). No significant difference was observed for freeze-thawed serum (P = 0.430). Conclusions: Plasma RNA is stable in uncentrifuged EDTA blood stored at 4 °C, but to obtain a stable serum RNA concentration, uncentrifuged clotted blood should be stored at 4 °C and processed within 6 h. A single freeze/thaw cycle produces no significant effect on the RNA concentration of plasma or serum. © 2002 American Association for Clinical Chemistry.
Persistent Identifierhttp://hdl.handle.net/10722/172809
ISSN
2015 Impact Factor: 7.457
2015 SCImago Journal Rankings: 2.472
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorTsui, NBYen_US
dc.contributor.authorNg, EKOen_US
dc.contributor.authorLo, YMDen_US
dc.date.accessioned2012-10-30T06:25:02Z-
dc.date.available2012-10-30T06:25:02Z-
dc.date.issued2002en_US
dc.identifier.citationClinical Chemistry, 2002, v. 48 n. 10, p. 1647-1653en_US
dc.identifier.issn0009-9147en_US
dc.identifier.urihttp://hdl.handle.net/10722/172809-
dc.description.abstractBackground: Circulating RNA in plasma/serum is an emerging field for noninvasive molecular diagnosis. Because RNA is widely thought to be labile in the circulation, we investigated the stability and various preanalytical factors that may affect RNA concentrations in blood specimens. Methods: Blood samples were collected from 65 healthy volunteers. The effects of two preanalytical variables Were studied: (a) time delay in processing of EDTA blood and clotted blood after venesection, and (b) freezing and thawing of plasma and serum. The lability of free added RNA in plasma was also investigated. Plasma/serum RNA was measured by a real-time quantitative reverse transcription-PCR assay for glyceraldehyde 3-phosphate dehydrogenase mRNA, whereas DNA was measured by a real-time quantitative PCR assay for the β-globin gene. Results: No significant difference was found for plasma RNA concentrations obtained from uncentrifuged EDTA blood that had been left at 4 °C for 0, 6, and 24 h (P = 0.182). On the other hand, the serum RNA concentrations increased significantly over 24 h when uncentrifuged clotted blood was stored at 4 °C (P <0.05). In comparison, >99% of the free added RNA could no longer be amplified after incubation in plasma for 15 s. Never-frozen plasma, freeze-thawed plasma, and thawed plasma left at room temperature for 1 h showed no significant differences in RNA concentration (P = 0:465). No significant difference was observed for freeze-thawed serum (P = 0.430). Conclusions: Plasma RNA is stable in uncentrifuged EDTA blood stored at 4 °C, but to obtain a stable serum RNA concentration, uncentrifuged clotted blood should be stored at 4 °C and processed within 6 h. A single freeze/thaw cycle produces no significant effect on the RNA concentration of plasma or serum. © 2002 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 Specimen Collectionen_US
dc.subject.meshDna - Blooden_US
dc.subject.meshDrug Stabilityen_US
dc.subject.meshGlobins - Geneticsen_US
dc.subject.meshGlyceraldehyde-3-Phosphate Dehydrogenases - Geneticsen_US
dc.subject.meshHumansen_US
dc.subject.meshPlasmaen_US
dc.subject.meshPolymerase Chain Reactionen_US
dc.subject.meshRna - Blooden_US
dc.subject.meshTime Factorsen_US
dc.titleStability of endogenous and added RNA in blood specimens, serum, and 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.pmid12324479-
dc.identifier.scopuseid_2-s2.0-0036794062en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0036794062&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume48en_US
dc.identifier.issue10en_US
dc.identifier.spage1647en_US
dc.identifier.epage1653en_US
dc.identifier.isiWOS:000178238100002-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridTsui, NBY=6602401748en_US
dc.identifier.scopusauthoridNg, EKO=21135553700en_US
dc.identifier.scopusauthoridLo, YMD=7401935391en_US

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