alexa A Novel Method for Mutation Analysis Using Genomic DNA

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A Novel Method for Mutation Analysis Using Genomic DNA

Tumor biopsies obtained from patients are often limited in size and availability, and the ability to perform multiple diagnostic assays depends on the quantity and quality of the tissue. Here we describe and evaluate a method for performing DNA-based mutational analyses after immunohistochemistry analysis has been performed, using a single tissue section.Results demonstrate that genomic DNA isolated from immunohistochemistry-stained and unstained formalin-fixed paraffin-embedded tissue sections are comparable in quality and are suitable for down-stream analysis using polymerase chain reaction based assays. We also found that the sensitivity and specificity in detecting hotspot mutations are comparable in both sources of genomic DNA. This study reports 100% concordance in detecting hotspot mutations in KRAS, BRAF, NRAS and PIK3CA using quantitative real-time polymerase chain reaction between stained and unstained formalin-fixed paraffin-embedded sections. Conclude that by using our novel approach, it is possible to perform immunohistochemistry staining followed by genomic analysis using a single 4-5 μm section of formalin-fixed paraffin-embedded tissue.


Citation: Desai R, Patel R, Amler L, Koeppen H, McCaffery I, et al. (2015) A Novel Method for Mutation Analysis Using Genomic DNA Obtained from Immunohistochemistry-Stained Sections. J Mol Biomark Diagn 6:228

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