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https://hdl.handle.net/2440/63738
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DC Field | Value | Language |
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dc.contributor.author | McMichael, G. | - |
dc.contributor.author | Highet, A. | - |
dc.contributor.author | Gibson, C. | - |
dc.contributor.author | Goldwater, P. | - |
dc.contributor.author | O'Callaghan, M. | - |
dc.contributor.author | Alvino, E. | - |
dc.contributor.author | MacLennan, A. | - |
dc.date.issued | 2011 | - |
dc.identifier.citation | Journal of Biomolecular Techniques, 2011; 22(1):5-9 | - |
dc.identifier.issn | 1524-0215 | - |
dc.identifier.issn | 1943-4731 | - |
dc.identifier.uri | http://hdl.handle.net/2440/63738 | - |
dc.description.abstract | Reliable 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.statementofresponsibility | Gai 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.iso | en | - |
dc.publisher | The Association of Biomolecular Resource Facilities | - |
dc.rights | Copyright © 2011 Association of Biomolecular Resource Facilities | - |
dc.source.uri | http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3059536/?report=abstract&tool=pmcentrez | - |
dc.subject | South Australian Cerebral Palsy Research Group | - |
dc.subject | Humans | - |
dc.subject | Cytomegalovirus | - |
dc.subject | Herpesvirus 4, Human | - |
dc.subject | Pregnancy Complications, Infectious | - |
dc.subject | Cytomegalovirus Infections | - |
dc.subject | DNA, Viral | - |
dc.subject | Neonatal Screening | - |
dc.subject | Blood Specimen Collection | - |
dc.subject | Viral Load | - |
dc.subject | Analysis of Variance | - |
dc.subject | Sensitivity and Specificity | - |
dc.subject | Retrospective Studies | - |
dc.subject | Polymerase Chain Reaction | - |
dc.subject | Pregnancy | - |
dc.subject | Infant, Newborn | - |
dc.subject | Female | - |
dc.subject | Analytic Sample Preparation Methods | - |
dc.title | Comparison of DNA extraction methods from small samples of newborn screening cards suitable for retrospective perinatal viral research | - |
dc.type | Journal article | - |
dc.contributor.organisation | South Australian Cerebral Palsy Research Group | - |
pubs.publication-status | Published | - |
dc.identifier.orcid | McMichael, G. [0000-0002-6811-5301] | - |
dc.identifier.orcid | Goldwater, P. [0000-0003-4822-8488] | - |
dc.identifier.orcid | O'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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