Please use this identifier to cite or link to this item: https://hdl.handle.net/2440/63738
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dc.contributor.authorMcMichael, G.-
dc.contributor.authorHighet, A.-
dc.contributor.authorGibson, C.-
dc.contributor.authorGoldwater, P.-
dc.contributor.authorO'Callaghan, M.-
dc.contributor.authorAlvino, E.-
dc.contributor.authorMacLennan, A.-
dc.date.issued2011-
dc.identifier.citationJournal of Biomolecular Techniques, 2011; 22(1):5-9-
dc.identifier.issn1524-0215-
dc.identifier.issn1943-4731-
dc.identifier.urihttp://hdl.handle.net/2440/63738-
dc.description.abstractReliable detection of viral DNA in stored newborn screening cards (NSC) would give important insight into possible silent infection during pregnancy and around birth. We sought a DNA extraction method with sufficient sensitivity to detect low copy numbers of viral DNA from small punch samples of NSC. Blank NSC were spotted with seronegative EDTA-blood and seropositive EBV EDTA-blood. DNA was extracted with commercial and noncommercial DNA extraction methods and quantified on a spectrofluorometer using a PicoGreen dsDNA quantification kit. Serial dilutions of purified viral DNA controls determined the sensitivity of the amplification protocol, and seropositive EBV EDTA-blood amplified by nested PCR (nPCR) validated the DNA extraction methods. There were considerable differences between the commercial and noncommercial DNA extraction methods (P=0.014; P=0.016). Commercial kits compared favorably, but the QIamp DNA micro kit with an added forensic filter step was marginally more sensitive. The mean DNA yield from this method was 3 ng/μl. The limit of detection was 10 viral genome copies in a 50-μl reaction. EBV nPCR detection in neat and 1:10 diluted DNA extracts could be replicated reliably. We conclude that the QIamp Micro DNA extraction method with the added forensic spin-filter step was suitable for retrospective DNA viral assays from NSC.-
dc.description.statementofresponsibilityGai L. McMichael, Amanda R. Highet, Catherine S. Gibson, Paul N. Goldwater, Michael E. O’Callaghan, Emily R. Alvino and Alastair H. MacLennan for the South Australian Cerebral Palsy Research Group-
dc.language.isoen-
dc.publisherThe Association of Biomolecular Resource Facilities-
dc.rightsCopyright © 2011 Association of Biomolecular Resource Facilities-
dc.source.urihttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC3059536/?report=abstract&tool=pmcentrez-
dc.subjectSouth Australian Cerebral Palsy Research Group-
dc.subjectHumans-
dc.subjectCytomegalovirus-
dc.subjectHerpesvirus 4, Human-
dc.subjectPregnancy Complications, Infectious-
dc.subjectCytomegalovirus Infections-
dc.subjectDNA, Viral-
dc.subjectNeonatal Screening-
dc.subjectBlood Specimen Collection-
dc.subjectViral Load-
dc.subjectAnalysis of Variance-
dc.subjectSensitivity and Specificity-
dc.subjectRetrospective Studies-
dc.subjectPolymerase Chain Reaction-
dc.subjectPregnancy-
dc.subjectInfant, Newborn-
dc.subjectFemale-
dc.subjectAnalytic Sample Preparation Methods-
dc.titleComparison of DNA extraction methods from small samples of newborn screening cards suitable for retrospective perinatal viral research-
dc.typeJournal article-
dc.contributor.organisationSouth Australian Cerebral Palsy Research Group-
pubs.publication-statusPublished-
dc.identifier.orcidMcMichael, G. [0000-0002-6811-5301]-
dc.identifier.orcidGoldwater, P. [0000-0003-4822-8488]-
dc.identifier.orcidO'Callaghan, M. [0000-0001-5038-5859] [0000-0002-8178-9714]-
Appears in Collections:Aurora harvest 5
Cerebral Palsy Research Group publications
Obstetrics and Gynaecology publications

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