microRNAs are small regulatory RNAs that are currently emerging as new biomarkers for cancer and other diseases. In order for biomarkers to be useful in clinical settings, they should be accurately and reliably detected in clinical samples such as formalin fixed paraffin embedded (FFPE) sections and blood serum or plasma. These types of samples represent a challenge in terms of microRNA quantification. A newly developed method for microRNA qPCR using Locked Nucleic Acid (LNA)-enhanced primers enables accurate and reproducible quantification of microRNAs in scarce clinical samples. Here we show that LNA-based microRNA qPCR enables biomarker screening using very low amounts of total RNA from FFPE samples and the results are compared to microarray analysis data. We also present evidence that the addition of a small carrier RNA prior to total RNA extraction, improves microRNA quantification in blood plasma and laser capture microdissected (LCM) sections of FFPE samples.

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.ymeth.2010.01.006DOI Listing

Publication Analysis

Top Keywords

microrna quantification
12
total rna
12
clinical samples
12
microrna qpcr
8
ffpe samples
8
samples
6
improved microrna
4
quantification
4
quantification total
4
rna
4

Similar Publications

Background: The content of circulating exosomes has been observed to be altered in response to changes in physiological and pathological conditions, and they are detectable in different human fluids such as blood. Studies focused on the quantification of Aβ and tau proteins, as molecules contained within exosomes, suggest that they are related with Alzheimer disease (AD) and frontotemporal dementia (FTD) development, demonstrated that plasma-derived exosome analysis is a good approach for searching for biomarkers in the development of dementia. Our aim is to identify new blood biomarkers to detect the AD or FTD in the Chilean population using machine learning based on exosomal miRNAs.

View Article and Find Full Text PDF

Extracellular vesicles (EVs) in cerebrospinal fluid (CSF) represent a valuable source of biomarkers for central nervous system (CNS) diseases, offering new pathways for diagnosis and monitoring. However, existing methods for isolating EVs from CSF often prove to be labor-intensive and reliant on specialized equipment, hindering their clinical application. In this study, we present a novel, clinically compatible method for isolating EVs from CSF.

View Article and Find Full Text PDF

Background: The diagnosis of malignant thyroid nodules is mainly based on the fine-needle aspiration biopsy (FNAB). To improve the detection of malignant nodules, different molecular tests have been developed. We present a new molecular signature based on altered miRNA expressions and specific mutations.

View Article and Find Full Text PDF

Dual-signal portable microRNA biosensor based on a photothermal/visual strategy induced by cascading amplification techniques and horseradish peroxidase.

Talanta

January 2025

Key Laboratory of Medicinal Chemistry and Molecular Diagnosis of the Ministry of Education, Key Laboratory of Analytical Science and Technology of Hebei Province, College of Chemistry and Materials Science, Hebei University, Baoding 071002, Hebei Province, PR China. Electronic address:

MicroRNAs (miRNAs) serve as potential biomarkers for many diseases such as cancer, neurodegenerative diseases and cardiovascular conditions. The portable and accurate detection of miRNA is of great significance for the early diagnosis, treatment optimization and prognostic evaluation of diseases. Herein, a photothermal/visual dual-mode assay for let-7a is developed utilizing oxidized 3, 3', 5, 5' - tetramethylbenzidine (oxTMB) as signal reporter.

View Article and Find Full Text PDF

Background: Extracellular vesicles (EVs) are essential for cell-to-cell communication because they transport functionally active molecules, including proteins, RNA, and lipids, from secretory cells to nearby or distant target cells. Seminal plasma contains a large number of EVs (sEVs) that are phenotypically heterogeneous. The aim of the present study was to identify the RNA species contained in two subsets of porcine sEVs of different sizes, namely small sEVs (S-sEVs) and large sEVs (L-sEVs).

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!