Cell-free DNA cfDNA , a burgeoning class of molecular biomarkers, has been extensively studied across a variety of biomedical fields. As a key component of liquid biopsy, cfDNA testing is gaining prominence in disease detection and management due to the convenience of sample collection and the abundant wealth of genetic information it provides.
However, the broader clinical application of cfDNA is currently impeded by a lack of standardization in the preanalytical procedures for cfDNA analysis. A number of fundamental challenges, including the selection of appropriate preanalytical procedures, prevention of short cfDNA fragment loss, and the validation of various cfDNA measurement methods, remain unaddressed. These existing hurdles lead to difficulties in comparing results and ensuring repeatability, thereby undermining the reliability of cfDNA analysis in clinical settings.
This review discusses the crucial preanalytical factors that influence cfDNA analysis outcomes, including sample collection, transportation, temporary storage, processing, extraction, quality control, and long-term storage. The review provides clarification on achievable consensus and offers an analysis of the current issues with the goal of standardizing preanalytical procedures for cfDNA analysis.
Subsequent research demonstrated elevated levels of cfDNA in patients with cancer Song et al. Originating from apoptotic or necrotic events and active release mechanisms in the presence of intracellular circulating nucleases Qi et al.
Due to the stability of cfDNA molecules in body fluids Polini et al. However, the progress of most related studies remains in the preliminary stage, likely due to the challenges presented by preanalytical variables. The journey from sample collection to cfDNA analysis is intricate and involves several steps, such as preparation, collection, transportation, temporary storage, processing, extraction, quality control, and long-term storage Figure 1. Each step involves numerous conditions or details, and the variables interact with each other.
