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Published on: July 12, 2022
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Simultaneous extraction and quantification of circulating mitochondrial and nuclear DNA using a single plasma sample
Sandeep Kumar Vishwakarma1, Nusrath Fathima1, Santosh K Tiwari1
1Central Laboratory for Stem Cell Research and Translational Medicine, Centre for Liver Research and Diagnostics, Deccan College of Medical Sciences, Kanchanbagh, Hyderabad, Telangana, India.
Mitochondrion
|December 12, 2022
Summary
Researchers developed a method to extract circulating mitochondrial DNA (cir-mtDNA) and nuclear DNA (cir-ncDNA) from plasma. This technique helps differentiate type 2 diabetes patients from healthy individuals, clarifying diagnostic roles.
Area of Science:
- Biochemistry
- Molecular Biology
- Clinical Diagnostics
Background:
- Variations in circulating mitochondrial DNA (cir-mtDNA) and nuclear DNA (cir-ncDNA) levels are observed across diseases.
- The diagnostic significance of these circulating DNA types requires a discriminative evaluation approach.
- Existing methods may not clearly distinguish the roles of cir-mtDNA versus cir-ncDNA in disease diagnostics.
Purpose of the Study:
- To establish an in-house process for co-extracting cir-mtDNA and cir-ncDNA from a single plasma sample.
- To assess the utility of this method in discriminating type 2 diabetes mellitus (T2DM) patients from healthy controls.
- To resolve disputes regarding the discriminative role of cir-mtDNA and cir-ncDNA and enhance their diagnostic value.
Main Methods:
- Development of a standardized in-house protocol for DNA extraction.
- Simultaneous isolation of both cir-mtDNA and cir-ncDNA from human plasma.
- Comparative analysis of extracted DNA levels between T2DM patients and healthy individuals.
Main Results:
- Successful co-extraction of cir-mtDNA and cir-ncDNA from individual plasma samples was achieved.
- The method demonstrated potential in differentiating T2DM patients from healthy controls.
- The study provides a basis for understanding the distinct diagnostic contributions of cir-mtDNA and cir-ncDNA.
Conclusions:
- The developed co-extraction method is a precise tool for analyzing circulating DNAs.
- This approach aids in distinguishing the diagnostic impact of cir-mtDNA from cir-ncDNA.
- The findings support the improved diagnostic utility of analyzing both DNA types concurrently.
