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Difficulties detecting miRNA-203 in human whole saliva by the use of PCR
Martin Lundegard1, Karin Nylander, Karin Danielsson
1Department of Odontology, Umeå University, SE-901 85 Umeå, Sweden, karin.danielsson@odont.umu.se.
Objectives:
Oral Lichen Planus (OLP) is a chronic disease of the oral mucosa, and according to the WHO also a pre malignant condition. Micro-RNAs are short non coding RNAs capable of regulating mRNA expression. MiRNA:scan be detected in tissue, blood and human whole saliva, HWS, and recently we have shown miR-203 to be up-regulated in tissue from OLP lesions.
Study Design:
In order to see whether mRNA as well asmiR-203 could be detected also in HWS, saliva from healthy controls and patients with OLP were analysed using two different PCR methods.
Results:
RESULTS showed low mRNA and miRNAlevels in general in HWS samples, making it hard to generate conclusive results.
Conclusions:
In order to make HWS a valuable source for different analyses, more sensitive PCR techniques capable of detecting very low levels of mRNAand miRNAas well as more efficient methods for extraction of RNA are needed.
Insights
Detecting oral lichen planus biomarkers in saliva is challenging due to low levels of micro-RNA (miRNA) and messenger RNA (mRNA). More sensitive techniques are needed to analyze human whole saliva (HWS) for diagnosing this pre-malignant condition.
Area of Science:
- Biochemistry
- Molecular Biology
- Oral Pathology
Background:
- Oral Lichen Planus (OLP) is a chronic oral mucosa disease with pre-malignant potential.
- Micro-RNAs (miRNAs) are regulatory molecules found in various bodily fluids, including human whole saliva (HWS).
- Previous research indicated elevated miR-203 levels in OLP tissue lesions.
Purpose of the Study:
- To investigate the detectability of mRNA and miR-203 in HWS from OLP patients and healthy controls.
- To assess the feasibility of using HWS as a diagnostic biomarker source for OLP.
Main Methods:
- Analysis of saliva samples from OLP patients and healthy individuals.
- Utilized two distinct Polymerase Chain Reaction (PCR) methods for detection.
- Quantified levels of specific mRNA and miR-203.
Main Results:
- Low concentrations of both mRNA and miRNA were generally observed in HWS samples.
- The low levels hindered the generation of conclusive analytical results.
- Detecting these biomarkers in HWS proved challenging with current methods.
Conclusions:
- Human whole saliva (HWS) currently presents limitations as a source for mRNA and miRNA analysis due to low analyte levels.
- Development of more sensitive PCR techniques is required for detecting low-abundance mRNA and miRNA.
- Improved RNA extraction methods are necessary to enhance the utility of HWS for diagnostic analyses.
