Determination of Xanthine Dehydrogenase mRNA by a Reverse Transcription-Coupled Competitive Quantitative Polymerase Chain Reaction Assay: Regulation in Rat Endothelial Cells by Hypoxia and Hyperoxia

1996 ◽  
Vol 335 (2) ◽  
pp. 377-380 ◽  
Author(s):  
Joseph J. Lanzillo ◽  
Feng-Sheng Yu ◽  
Joanne Stevens ◽  
Paul M. Hassoun
Keyword(s):  
Polymerase Chain Reaction ◽  
Endothelial Cells ◽  
Reverse Transcription ◽  
Polymerase Chain Reaction Assay ◽  
Quantitative Polymerase Chain Reaction ◽  
Xanthine Dehydrogenase ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals Technical Validation of a Reverse-Transcription Quantitative Polymerase Chain Reaction In Vitro Diagnostic Test for the Determination of MiR-31-3p Expression Levels in Formalin-Fixed Paraffin-Embedded Metastatic Colorectal Cancer Tumor Specimens

2018 ◽  
Vol 13 ◽  
pp. 117727191876335 ◽  
Author(s):  
Lucas Ramon ◽  
Catherine David ◽  
Karine Fontaine ◽  
Elodie Lallet ◽  
Charles Marcaillou ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Polymerase Chain ◽  
Formalin Fixed

MiR-31-3p expression has been shown to be a predictive biomarker for response to anti-epithelial growth factor receptor therapy in patients with RAS wild-type metastatic colorectal cancer (mCRC). To aid in the quantification of miR-31-3p expression in formalin-fixed paraffin-embedded (FFPE) primary tumor samples from patients with mCRC, a reverse-transcription quantitative polymerase chain reaction (RT-qPCR) assay was developed and validated. Assay development included the identification of a microRNA reference standard and the determination of an appropriate relative quantification cutoff for differentiating low versus high miR-31-3p expression. Sample specimens for the validation studies included both FFPE slides and shavings. Polymerase chain reaction (PCR) efficiency and linearity, analytical sensitivity and specificity, assay robustness, reproducibility, and accuracy were demonstrated across a number of test conditions and differing quantitative PCR platforms. The data from this study provide evidence as to the feasibility of quantifying the expression of miR-31-3p from FFPE tumor tissue using a standardized RT-qPCR assay.

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