Ratio-Based Method To Identify True Biomarkers by Normalizing Circulating ncRNA Sequencing and Quantitative PCR Data. Academic Article uri icon

Overview

abstract

  • Recent studies have indicated that circulating noncoding RNAs (ncRNAs) such as miRNAs are stable biomarkers for the diagnosis and prognosis of human diseases. However, due to low concentrations of circulating ncRNAs in blood, data normalization in plasma/serum ncRNA experiments using next-generation sequencing and quantitative real time RT-qPCR is a challenge. We found that the current normalization methods based on synthetic external spiked-in controls or published endogenous miRNA controls are inappropriate as they are not stably expressed and therefore fail to reliably detect differentially expressed ncRNAs. Using the alternative of individual ncRNAs as biomarkers, we considered a ratio-based normalization method calculated taking the ratio of any two ncRNAs in the same sample and used the resulting ratios as biomarkers. We mathematically verified the method to be independent of spiked-in and internal controls, and more robust than existing reference control based normalization methods to identify differentially expressed ncRNAs as potential biomarkers for human diseases. Thus, the ratio-based method can solve the difficult normalization problem for circuiting ncRNA data to identify reliable biomarkers to meet real clinical practice.

publication date

  • April 30, 2019

Research

keywords

  • Biomarkers, Tumor
  • Circulating MicroRNA
  • Real-Time Polymerase Chain Reaction
  • Sequence Analysis, RNA

Identity

PubMed Central ID

  • PMC6884007

Scopus Document Identifier

  • 85065872901

Digital Object Identifier (DOI)

  • 10.1155/2016/1869082

PubMed ID

  • 31002238

Additional Document Info

volume

  • 91

issue

  • 10