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Article: Effects of blood-processing protocols on fetal and total DNA quantification in maternal plasma

TitleEffects of blood-processing protocols on fetal and total DNA quantification in maternal plasma
Authors
Issue Date2001
PublisherAmerican Association for Clinical Chemistry, Inc. The Journal's web site is located at http://www.clinchem.org
Citation
Clinical Chemistry, 2001, v. 47 n. 9, p. 1607-1613 How to Cite?
AbstractBackground: Recently, apoptotic cells have been found in plasma obtained by centrifugation of blood from pregnant women, raising the question of what constitutes plasma and whether plasma is truly cell free. We compared the effects of different blood-processing protocols on the quantification, DNA composition, and day-to-day fluctuation of fetal and total DNA in maternal plasma. Methods: Blood samples were collected from healthy pregnant women. The blood sample from each individual was simultaneously processed by different means, including the following: Percoll separation, centrifugation, microcentrifugation, and filtration. The resulting plasma aliquots were subjected to real-time quantitative amplification of the β-globin (for total DNA) and SRY (for fetal DNA) genes. The differences in the β-globin and SRY DNA concentrations and the degree of variation between the various plasma aliquots were assessed statistically. Results: Different protocols of blood processing significantly affected the quantification and the day-to-day fluctuation of total (P <0.001), but not fetal (quantification, P = 0.336; fluctuation, P = 0.206), DNA in maternal plasma. The quantitative difference could be attributed to the fact that efficacies of different protocols for generating cell-free plasma vary. Processing blood samples by centrifugation followed by filtration or microcentrifugation is effective in producing cell-free plasma. Conclusions: Standardization in plasma-processing protocols is needed for maternal plasma DNA analysis, especially for quantification of total DNA in maternal plasma. Such preanalytic factors may also affect other applications of plasma DNA analysis. © 2001 American Association for Clinical Chemistry.
Persistent Identifierhttp://hdl.handle.net/10722/179769
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.authorPoon, LLMen_US
dc.contributor.authorLau, TKen_US
dc.contributor.authorLeung, TNen_US
dc.contributor.authorWong, EMCen_US
dc.contributor.authorLo, YMDen_US
dc.date.accessioned2012-12-19T10:04:27Z-
dc.date.available2012-12-19T10:04:27Z-
dc.date.issued2001en_US
dc.identifier.citationClinical Chemistry, 2001, v. 47 n. 9, p. 1607-1613en_US
dc.identifier.issn0009-9147en_US
dc.identifier.urihttp://hdl.handle.net/10722/179769-
dc.description.abstractBackground: Recently, apoptotic cells have been found in plasma obtained by centrifugation of blood from pregnant women, raising the question of what constitutes plasma and whether plasma is truly cell free. We compared the effects of different blood-processing protocols on the quantification, DNA composition, and day-to-day fluctuation of fetal and total DNA in maternal plasma. Methods: Blood samples were collected from healthy pregnant women. The blood sample from each individual was simultaneously processed by different means, including the following: Percoll separation, centrifugation, microcentrifugation, and filtration. The resulting plasma aliquots were subjected to real-time quantitative amplification of the β-globin (for total DNA) and SRY (for fetal DNA) genes. The differences in the β-globin and SRY DNA concentrations and the degree of variation between the various plasma aliquots were assessed statistically. Results: Different protocols of blood processing significantly affected the quantification and the day-to-day fluctuation of total (P <0.001), but not fetal (quantification, P = 0.336; fluctuation, P = 0.206), DNA in maternal plasma. The quantitative difference could be attributed to the fact that efficacies of different protocols for generating cell-free plasma vary. Processing blood samples by centrifugation followed by filtration or microcentrifugation is effective in producing cell-free plasma. Conclusions: Standardization in plasma-processing protocols is needed for maternal plasma DNA analysis, especially for quantification of total DNA in maternal plasma. Such preanalytic factors may also affect other applications of plasma DNA analysis. © 2001 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 Collection - Methodsen_US
dc.subject.meshDna - Blood - Isolation & Purificationen_US
dc.subject.meshFemaleen_US
dc.subject.meshFetal Blood - Chemistryen_US
dc.subject.meshHumansen_US
dc.subject.meshPolymerase Chain Reactionen_US
dc.subject.meshPregnancy - Blooden_US
dc.titleEffects of blood-processing protocols on fetal and total DNA quantification in maternal plasmaen_US
dc.typeArticleen_US
dc.identifier.emailPoon, LLM: llmpoon@hkucc.hku.hken_US
dc.identifier.authorityPoon, LLM=rp00484en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.pmid11514393-
dc.identifier.scopuseid_2-s2.0-0034872431en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0034872431&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume47en_US
dc.identifier.issue9en_US
dc.identifier.spage1607en_US
dc.identifier.epage1613en_US
dc.identifier.isiWOS:000170636600004-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridChiu, RWK=7103038413en_US
dc.identifier.scopusauthoridPoon, LLM=7005441747en_US
dc.identifier.scopusauthoridLau, TK=24491963900en_US
dc.identifier.scopusauthoridLeung, TN=7202110927en_US
dc.identifier.scopusauthoridWong, EMC=13409091100en_US
dc.identifier.scopusauthoridLo, YMD=7401935391en_US

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