Bacterial profile-based body fluid identification using a machine learning approach.

Abstract

Identifying the origins of biological traces is critical for the reconstruction of crime scenes in forensic investigations. Traditional methods for body fluid identification rely on chemical, enzymatic, immunological, and spectroscopic techniques, which can be sample-consuming and depend on simple color-change reactions. However, these methods have limitations when residual samples are insufficient after DNA extraction. This study aimed to develop a method for body fluid identification by leveraging bacterial DNA profiling to overcome the limitations of the conventional approaches. Bacterial profiles were determined by sequencing the hypervariable region of the 16S rRNA gene,using DNA metabarcoding of evidence collected from criminal cases. Amplicon sequence variants (ASVs) were analyzed to identify significant microbial patterns in different body fluid samples. The bacterial profile-based method demonstrated high discriminatory power with a machine learning model trained using the naïve Bayes algorithm, achieving an accuracy of over 98% in classifying samples into one of four body fluid types: blood, saliva, vaginal secretion, and mixture traces of vaginal secretions and semen. Bacterial profiling enhances the accuracy and robustness of body fluid identification in forensic analysis, providing a valuable alternative to traditional methods by utilizing DNA and microbial community data despite the uncontrollable conditions. This approach offers significant improvements in the classification accuracy and practical applicability in forensic investigations.