Controls to validate plasma samples for cell free DNA quantification.

Area of Science:

• Biochemistry
• Molecular Biology
• Clinical Diagnostics

Background:

• Cell-free DNA (cfDNA) in serum and plasma shows promise as a tumor marker for cancer treatment selection, monitoring, and follow-up.
• Diverging results in current literature are attributed to methodological inconsistencies and a lack of standardization, hindering routine clinical application.
• Existing biological insights from cfDNA analysis have not yet been integrated into standard clinical practice.

Purpose of the Study:

• To present a novel method for external standardization of cfDNA analysis using non-human DNA fragments.
• To introduce a technique for controlling DNA loss during sample preparation and measurement.
• To develop a method for accounting for normal lymphocyte DNA admixture using B-cell immunoglobulin gene rearrangements.

Main Methods:

• External standardization by spiking samples with non-human DNA fragments.
• Utilizing unique immunoglobulin gene rearrangements in B-cells to identify and control for normal lymphocyte DNA.
• Implementing these methods to assess cfDNA levels in serum and plasma samples.

Main Results:

• The proposed approach enhances the quality of cfDNA analysis.
• The method effectively lowers the risk of falsely elevated cfDNA values.
• Improved accuracy in cfDNA measurements was demonstrated.

Conclusions:

• The developed method offers improved accuracy for cfDNA measurements.
• This technique is easily incorporated into existing cfDNA analysis technologies.
• The standardization approach facilitates the routine clinical use of cfDNA as a tumor marker.