Evaluation and selection of a non-PCR based technology for improved gene expression profiling from clinical formalin-fixed, paraffin-embedded samples

Author:

Qi Zhenhao1,Wang Lisu1,He Aiqing2,Ma-Edmonds Manling2,Cogswell John1

Affiliation:

1. Clinical Translational Technologies & Operations, Bristol-Myers Squibb, Route 206 & Province Line Road, Princeton, NJ 08543, USA

2. Genetically Defined Diseases & Genomics, Bristol-Myers Squibb, Route 206 & Province Line Road, Princeton, NJ 08543, USA

Abstract

Aim: Formalin-fixed, paraffin-embedded (FFPE) clinical tissue samples have the potential to provide valuable gene-expression data for the development of cancer biomarkers. However, FFPE RNA is extensively fragmented, presenting a significant challenge for reliably detecting gene expression using traditional qPCR methods. Results: We evaluated three novel methodologies along with the traditional qPCR method for their ability to detect Notch pathway gene expression in colorectal cancer FFPE tissue RNAs. We found that quantitative nuclease protection assay-detected gene expression in high-quality RNAs as sensitively as qPCR, and consistently detected mRNAs in highly fragmented FFPE tissue RNAs. Conclusion: Quantitative nuclease protection assay represents an improved methodology for detecting gene expression in FFPE tissue and has the potential to advance the development of cancer biomarkers.

Publisher

Future Science Ltd

Subject

Medical Laboratory Technology,Clinical Biochemistry,General Pharmacology, Toxicology and Pharmaceutics,General Medicine,Analytical Chemistry

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