Forensic Body Fluid RNA Typing: Evaluating the Roles of CE and dPCR Technologies

Area of Science:

• Forensic Science
• Molecular Biology
• Biochemistry

Background:

• RNA-based methods are crucial for body fluid identification in forensics, offering tissue specificity and information beyond DNA.
• Capillary electrophoresis (CE) is the traditional standard for RNA analysis in forensic labs, known for multiplexing.
• Digital PCR (dPCR) offers high sensitivity and absolute quantification, especially for degraded or low-template forensic samples.

Purpose of the Study:

• To comparatively evaluate capillary electrophoresis (CE) and digital PCR (dPCR) workflows for RNA-based body fluid identification.
• To assess the analytical performance and forensic applicability of both CE and dPCR platforms.
• To highlight the strengths and limitations of each technology for forensic casework.

Main Methods:

• Comparative analysis of CE- and dPCR-based workflows.
• Evaluation of key parameters: sensitivity, specificity, multiplexing capacity, RNA degradation tolerance, and interpretation.
• Review of existing literature on RNA-based body fluid identification techniques.

Main Results:

• CE excels in multiplex assay design and routine forensic implementation due to established infrastructure and throughput.
• dPCR demonstrates superior sensitivity and quantitative precision, particularly for challenging forensic samples.
• dPCR's current limitations include reduced multiplexing capacity and higher costs compared to CE.

Conclusions:

• CE and dPCR are complementary rather than competing technologies for RNA-based body fluid identification.
• Combined or context-dependent use of CE and dPCR can enhance the robustness of forensic analysis.
• Further validation and standardization are necessary for broader dPCR implementation in routine forensic workflows.