Female sample screening using colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP)...

Area of Science:

• Forensic Science
• Molecular Biology
• Genetics

Background:

• Effective DNA typing workflows rely on accurate forensic sample screening.
• Detecting sex-specific markers is crucial for forensic analysis.
• Current DNA-based methods struggle to confirm female contributions in forensic samples.

Purpose of the Study:

• To develop a novel assay for the specific screening of female forensic samples.
• To address the limitations of existing DNA-based methods for detecting female DNA.
• To provide a rapid and simple tool for forensic casework.

Main Methods:

• Development of a colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay.
• Targeting the X-inactive specific transcript (XIST) long non-coding RNA for detection.
• Assay operation at 65°C for 30 minutes with visual pH indicator dye readout.

Main Results:

• The assay demonstrated a detection limit of approximately 0.5 µL of blood.
• Successful detection of XIST RNA in mixed body fluids and mock forensic samples.
• Results were discernible through a simple color change, indicating assay functionality.

Conclusions:

• The developed RT-LAMP assay provides a rapid and simple strategy for screening female samples.
• The assay shows potential applicability for real-world forensic casework.
• This method enhances the ability to confirm female contributions in forensic investigations.