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dc.contributor.authorThorogate, R
dc.contributor.authorMoreira, JCS
dc.contributor.authorJickells, S
dc.contributor.authorMiele, MMP
dc.contributor.authorDaniel, B
dc.date.accessioned2018-08-30T13:42:13Z
dc.date.issued2008-09
dc.identifier4
dc.identifier.citationFORENSIC SCIENCE INTERNATIONAL-GENETICS, 2008, 2 pp. 363 - 371
dc.identifier.issn1872-4973
dc.identifier.urihttps://repository.icr.ac.uk/handle/internal/2442
dc.identifier.eissn1878-0326
dc.identifier.doi10.1016/j.fsigen.2008.06.003
dc.description.abstractFull DNA profiles can be generated from just a few cells; however these profiles can be contaminated from other cell types present at the crime scene. We report here on the development of an immunofluorescent technique to spatially locate human-specific blood in situ and also on the ability of this technique to detect individual leukocytes and the DNA contained within them. Four monoclonal Mouse anti-human antibodies were evaluated; anti-glycophorin A to detect erythrocytes and anti-CD45, anti-myeloperoxidase (MPO) and anti-histone H1 to detect the nucleated leukocytes. Each antibody was labeled with either Alexa Fluor 488 or 568 for direct application to blood smears which allowed the Simultaneous detection of erythrocytes and leukocytes. Furthermore, because histories are DNA binding proteins, the application of anti-histone H1 allowed the detection of DNA within a blood smear. Importantly it was found that full DNA profiles could be achieved after using this method with similar peak area ratios compared to untreated cells. The fluorescent antibodies were found to be human-specific with the exception of anti-histone H1 due to its conserved sequence. However, used in combination with anti-CD45 or anti-MPO the location of DNA from human-specific leukocytes could be detected. The technique was also tested on older blood stains and was still found to be sensitive and cell-specific after 4 months. Following the optimization of the methodology, the fluorescent antibodies were applied to short lengths of black cotton fibres covered with human blood spots. Although the background fluorescence from the cotton was found to be high, erythrocytes and even individual leukocytes could easily be detected, indicating that this technique could be used to detect extremely minute amounts of blood. Used in combination with laser capture microdissection (LCM), this method could be used to pick off individual leukocytes for LCN DNA techniques. (C) 2008 Elsevier Ireland Ltd. All rights reserved.
dc.format.extent363 - 371
dc.languageeng
dc.language.isoeng
dc.publisherELSEVIER IRELAND LTD
dc.titleA novel fluorescence-based method in forensic science for the detection of blood in situ
dc.typeJournal Article
rioxxterms.versionofrecord10.1016/j.fsigen.2008.06.003
rioxxterms.licenseref.startdate2008-09
rioxxterms.typeJournal Article/Review
dc.relation.isPartOfFORENSIC SCIENCE INTERNATIONAL-GENETICS
pubs.notesaffiliation: Thorogate, R (Reprint Author), Kings Coll London, Dept Forens Sci & Drug Monitoring, Franklin Wilkins Bldg,150 Stamford St, London SE1 9NH, England. Thorogate, Richard; Jickells, Sue; Miele, Margherita M. P.; Daniel, Barbara, Kings Coll London, Dept Forens Sci & Drug Monitoring, London SE1 9NH, England. Moreira, Joana C. S., Inst Canc Res, Sutton SM2 5NG, Surrey, England. keywords: Antibody; Blood; Fixation; Fluorophores; Immunofluorescence; Microscopy keywords-plus: DNA; SEQUENCE; HISTONE; MYELOPEROXIDASE; GLYCOPHORIN; ANTIBODIES; SUBTYPES; TESTS; PCR research-areas: Genetics & Heredity; Legal Medicine web-of-science-categories: Genetics & Heredity; Medicine, Legal author-email: [email protected] funding-acknowledgement: UK Engineering and Physical Sciences Research Council (EPSRC); University of Padua, Italy; Engineering and Physical Sciences Research Council [EP/D041082/1] funding-text: The authors wish to thank the UK Engineering and Physical Sciences Research Council (EPSRC) ‘Think Crime’ initiative for funding this project and the EU Soctrates Erasmus programme for funding Margherita Miele from the University of Padua, Italy. We also thank Ignacio Quinones, King’s College London, for his valuable assistance with the DNA profiling. number-of-cited-references: 30 times-cited: 16 usage-count-last-180-days: 2 usage-count-since-2013: 13 journal-iso: Forensic Sci. Int.-Genet. doc-delivery-number: 380XX unique-id: ISI:000261500500016 da: 2018-08-30
pubs.notesNot known
pubs.organisational-group/ICR
pubs.organisational-group/ICR
pubs.volume2
pubs.embargo.termsNot known
dc.contributor.icrauthorMoreira, Joanaen


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