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Molecular analysis of H&E - and Papanicolau-stained samples: systematic review

2020-05Journal article Restricted access
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dc.contributor.authorPote, Alexandra
dc.contributor.authorBoghenco, Otília
dc.contributor.authorMarques-Ramos, Ana
dc.date.accessioned2020-05-27T19:33:07Z
dc.date.available2020-05-27T19:33:07Z
dc.date.issued2020-05
dc.description.abstractMolecular pathology allows the identification of causative agents in infectious diseases and the detection of biomarkers important for prediction of disease susceptibility, diagnosis and personalized therapy. Accordingly, nucleic acid-based methods have gained a special role in clinical laboratories particularly to evaluate solid and hematological tumors. Extraction of nucleic acids is commonly performed in microdissected formalin-fixed paraffin-embedded (FFPE) or cytological samples that had been previously evaluated through the use of hematoxylin and eosin (H&E) or Papanicolau (Pap) stains, respectively. Although the effect of both stains on nucleic acids integrity has been explored by several authors, the results are not consistent and require further examination. Accordingly, the goal of this review was to assess the influence of H&E and Pap stains on DNA and RNA integrity and to address the mechanism by which each staining compromises molecular based-analysis. The analyzed studies demonstrate that H&E- and Pap-staining result in low DNA recovery and some degree of DNA fragmentation. Additionally, it is concluded that hemalum inhibits PCR by interfering with DNA extraction, preventing DNA polymerase attachment and possibly by rescuing divalent cations. Accordingly, proper sample purification and adjustment of PCR conditions are of key importance to achieve satisfactory results by PCR in H&E- and Pap-stained samples. Furthermore, although H&E results in RNA fragmentation, it is possible to perform expression analysis in H&E-stained frozen sections, using RNase-free conditions, low amounts of hematoxylin and a rapid protocol from sample collection to RNA analysis. It The effect of Pap-staining on RNA integrity remains to be determined.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationPote A, Boghenco O, Marques-Ramos A. Molecular analysis of H&E – And Papanicolau-stained samples: systematic review. Histochem Cell Biol. 2020;154:7-20.pt_PT
dc.identifier.doi10.1007/s00418-020-01882-wpt_PT
dc.identifier.urihttp://hdl.handle.net/10400.21/11741
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherSpringerpt_PT
dc.relation.publisherversionhttps://link.springer.com/article/10.1007/s00418-020-01882-wpt_PT
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/pt_PT
dc.subjectDNA integritypt_PT
dc.subjectHematoxylin and eosinpt_PT
dc.subjectH&e-stained samplespt_PT
dc.subjectPapanicolau-stained samplespt_PT
dc.subjectMolecular analysispt_PT
dc.subjectRNA integritypt_PT
dc.titleMolecular analysis of H&E - and Papanicolau-stained samples: systematic reviewpt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage20pt_PT
oaire.citation.startPage7pt_PT
oaire.citation.titleHistochemistry and Cell Biologypt_PT
oaire.citation.volume154pt_PT
person.familyNameMarques-Ramos
person.givenNameAna
person.identifier.ciencia-id0617-3550-881E
person.identifier.orcid0000-0003-2655-8806
rcaap.rightsrestrictedAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublication0b8d516d-bf23-4db0-8224-d33e28d309ac
relation.isAuthorOfPublication.latestForDiscovery0b8d516d-bf23-4db0-8224-d33e28d309ac

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