RT Journal Article
SR Electronic
T1 Nucleic acid sequence-based amplification in formalin-fixed and paraffin-embedded breast-cancer tissues
JF Journal of Clinical Pathology
JO J Clin Pathol
FD BMJ Publishing Group Ltd and Association of Clinical Pathologists
SP 1071
OP 1076
DO 10.1136/jcp.2010.078766
VO 63
IS 12
A1 Riehle, Ulrike
A1 Mader, Andreas
A1 Brandstetter, Thomas
A1 Rühe, Jürgen
A1 zur Hausen, Axel
A1 Stickeler, Elmar
YR 2010
UL http://jcp.bmj.com/content/63/12/1071.abstract
AB 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.