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Nanopore-based detection of periodontitis biomarker miR31 in saliva samples
Pearl Arora1, Haiyan Zheng2, Sathishkumar Munusamy2
1Department of Chemistry, Illinois Institute of Technology, Chicago, Illinois, USA.
Abstract:
MicroRNAs (miRNAs) play important roles in posttranscriptional gene regulation. Aberrations in the miRNA levels have been the cause behind various diseases, including periodontitis. Therefore, sensitive, specific, and accurate detection of disease-associated miRNAs is vital to early diagnosis and can facilitate inhibitor screening and drug design. In this study, we developed a label-free, real-time sensing method for the detection of miR31, which has been frequently linked to periodontitis, using an engineered protein nanopore and in the presence of a complementary ssDNA as a molecular probe. Our method is rapid and highly sensitive with nanomolar concentration of miR31 that could be determined in minutes. Furthermore, our sensor showed high selectivity toward the target miR31 sequence even in the presence of interfering nucleic acids. In addition, artificial saliva and human saliva samples were successfully analyzed. Our developed nanopore sensing platform could be used to detect other miRNAs and offers a potential application for the clinical diagnosis of disease biomarkers.
Insights
We developed a novel nanopore sensor for rapid and sensitive detection of microRNAs (miRNAs) linked to periodontitis. This technology shows promise for early disease diagnosis and biomarker discovery.
Area of Science:
- Biotechnology
- Molecular Biology
- Nanotechnology
Background:
- MicroRNAs (miRNAs) are crucial in gene regulation, and their altered levels are implicated in diseases like periodontitis.
- Accurate detection of disease-associated miRNAs is essential for early diagnosis, inhibitor screening, and drug development.
Purpose of the Study:
- To develop a label-free, real-time sensing method for detecting miR31, a microRNA associated with periodontitis.
- To demonstrate the sensor's sensitivity, specificity, and applicability in biological samples.
Main Methods:
- Utilized an engineered protein nanopore system with a complementary single-stranded DNA (ssDNA) molecular probe.
- Employed a label-free, real-time detection approach for miR31 quantification.
Main Results:
- Achieved rapid detection of miR31 at nanomolar concentrations within minutes.
- Demonstrated high selectivity for the target miR31 sequence, even with interfering nucleic acids present.
- Successfully analyzed artificial and human saliva samples, validating the sensor's practical utility.
Conclusions:
- The developed nanopore sensing platform offers a sensitive and specific method for miR31 detection.
- This technology has potential applications in the clinical diagnosis of periodontitis and other diseases through biomarker detection.
