Optimized RT-qPCR and a novel normalization method for validating circulating miRNA biomarkers in ageing-related diseases.

ABSTRACT: Circulating miRNAs have potential as minimally invasive biomarkers for diagnosing various diseases, including ageing-related disorders such as Alzheimer's disease (AD). However, the lack of standardization in the common analysis method, RT-qPCR, and specifically in the normalization step, has resulted in inconsistent data across studies, hindering miRNA clinical implementation as well as basic research. To address this issue, this study proposes an optimized protocol for key steps in miRNA profiling, which incorporates absorbance-based haemolysis detection for assessing sample quality, double spike-in controls for miRNA isolation and reverse transcription, and the use of 7 stable normalizers verified in an aging population, including healthy subjects and individuals at different stages of

SUBMITTER: Want A 

PROVIDER: S-EPMC10682428 | biostudies-literature | 2023 Nov

REPOSITORIES: biostudies-literature