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Nucleic acid sequence-based amplification in formalin-fixed and paraffin-embedded breast-cancer tissues.
Riehle, Ulrike; Mader, Andreas; Brandstetter, Thomas; Rühe, Jürgen; zur Hausen, Axel; Stickeler, Elmar.
Afiliação
  • Riehle U; Department of Gynecology and Obstetrics, University Hospital Freiburg, Freiburg, Germany.
J Clin Pathol ; 63(12): 1071-6, 2010 Dec.
Article em En | MEDLINE | ID: mdl-20974626
AIM: To evaluate the nucleic acid sequence-based amplification (NASBA) technique to amplify mRNA isolated from formalin-fixed and paraffin-embedded (FFPE) breast-cancer tissues. METHODS: RNA was extracted from archived, 10-year-old FFPE tissues, and selected genes, namely ribosomal protein S18 (RPS18), epidermal growth factor receptor 2 (HER2), estrogen receptor alpha (ERα), Y box binding protein (YBX-1), matrix metallopeptidase 11 (MMP11), caspase 8 (CASP8) and superoxide dismutase 2 (SOD2), were amplified by NASBA. RESULTS: Despite strong degradation of the template, RNA amplification of all tested genes resulted in strong hybridisation signals. Sensitivity tests showed that the RPS18 NASBA assay was more sensitive than real-time RT-PCR used as a reference method. The sensitivity of the HER2, ERα, MMP11, YBX1, CASP8 and SOD2 NASBA assay was comparable with RT-PCR targeted to the respective genes. CONCLUSIONS: The results obtained indicate that NASBA is suitable to amplify with high specificity and sensitivity, even strongly degraded RNA isolated from FFPE tissues, and therefore can complement already-existing amplification techniques such as RT-PCR for analysis of such tissues.
Assuntos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Neoplasias da Mama / Replicação de Sequência Autossustentável / Genes Neoplásicos Tipo de estudo: Evaluation_studies Limite: Female / Humans Idioma: En Revista: J Clin Pathol Ano de publicação: 2010 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Neoplasias da Mama / Replicação de Sequência Autossustentável / Genes Neoplásicos Tipo de estudo: Evaluation_studies Limite: Female / Humans Idioma: En Revista: J Clin Pathol Ano de publicação: 2010 Tipo de documento: Article