A multiplex two-color real-time PCR method for quality-controlled molecular diagnostic testing of FFPE samples.

ABSTRACT:

Background

Reverse transcription quantitative real-time PCR (RT-qPCR) tests support personalized cancer treatment through more clinically meaningful diagnosis. However, samples obtained through standard clinical pathology procedures are formalin-fixed, paraffin-embedded (FFPE) and yield small samples with low integrity RNA containing PCR interfering substances. RT-qPCR tests able to assess FFPE samples with quality control and inter-laboratory reproducibility are needed.

Methods

We developed an RT-qPCR method by which 1) each gene was measured relative to a known number of its respective competitive internal standard molecules to control for interfering substances, 2) two-color fluorometric hydrolysis probes enabled analysis on a real-time platform, 3) external standards con

SUBMITTER: Yeo J 

PROVIDER: S-EPMC3931751 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature