DNA methylation-based assay and Random Forest classification model for identification of biological materials

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Key Points

The aim is to develop an assay for identifying biological materials based on DNA methylation patterns.
Developed a multiplex assay targeting 13 differentially methylated DNA positions (DMPs).
Utilized combined bisulfite conversion and SNaPshot analysis.
Constructed two Random Forest models for classification: RF1 for single-component samples and RF2 for mixtures.
Evaluated specificity and uncovered overlapping DNA methylation profiles.
RF1 achieved 92.7% accuracy for identifying single-component samples.
DNA input for analysis can range from 0.25 ng to 10 ng depending on the material.
Strong correlation exists between DNA methylation levels and two-component mixture proportions.
RF2 demonstrated promising performance for identifying unbalanced mixed samples.

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Develops an assay and classification model to identify biological materials, indicating its importance in forensic investigations.

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Schmelzer et al. (2026) studied this question.

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