Forensic DNA Recovery from FFPE Tissue Using the Maxwell^® RSC Xcelerate Kit: Optimization, Challenges, and...

Area of Science:

• Forensic science
• Molecular biology
• Genetics

Background:

• Archival formalin-fixed, paraffin-embedded (FFPE) tissues pose challenges for DNA profiling due to formalin-induced damage.
• Reliable DNA profiles are crucial for forensic investigations and medical evaluations, especially in cases involving cancer diagnosis or treatment.
• Retrospective molecular analyses of FFPE samples can provide critical evidence but are hindered by DNA quality issues.

Purpose of the Study:

• To evaluate the Maxwell® RSC Xcelerate DNA FFPE Kit for DNA profiling from endometrial cancer FFPE tissues.
• To identify limitations in generating DNA profiles from archival FFPE samples using this kit.

Main Methods:

• Analysis of archival endometrial cancer FFPE blocks using the Maxwell® RSC Xcelerate DNA FFPE Kit.
• Assessment of DNA yield, purity, and degradation using real-time PCR.
• Short tandem repeat (STR) profiling with forensic genotyping kits to evaluate profile completeness and reliability.

Main Results:

• The kit demonstrated efficient DNA recovery with low degradation indices from FFPE samples.
• Despite good DNA yield, complete STR profiles were frequently unsuccessful, showing allele dropout and imbalance.
• The quality of obtained DNA profiles was often insufficient for reliable forensic interpretation.

Conclusions:

• Generating complete and interpretable STR profiles from FFPE tissues remains a significant challenge, even with advanced extraction kits.
• DNA fragmentation and fixation artifacts in FFPE samples impair amplification efficiency and limit the evidentiary value of STR analysis.
• Further improvements in extraction protocols and interpretative strategies are needed to enhance DNA profiling reliability from archival FFPE material.