Rapid and simple sex determination method from dental pulp by loop-mediated isothermal amplification.

Area of Science:

• Forensic Science
• Molecular Biology
• Genetics

Background:

• Accurate sex determination from biological samples is crucial in forensic investigations.
• Dental pulp DNA offers a viable source for genetic analysis, even from aged samples.
• Existing methods for DNA sex determination can be time-consuming.

Purpose of the Study:

• To evaluate the efficacy of loop-mediated isothermal amplification (LAMP) for rapid sex determination using dental pulp DNA.
• To optimize primer design for the amelogenin locus to enhance speed and accuracy.
• To assess the applicability of the LAMP method on DNA from teeth stored for extended periods.

Main Methods:

• DNA was extracted from 32 dental pulp samples from permanent teeth aged 1-25 years post-extraction.
• A loop-mediated isothermal amplification (LAMP) assay targeting the amelogenin locus was developed.
• Four specific primers and loop primers were designed for rapid amplification.
• Real-time turbidimetry and visual inspection were used for detection of amplified products.

Main Results:

• Sex determination was achieved within approximately 32-34 minutes using the LAMP method.
• The inclusion of loop primers reduced the overall analysis time by half.
• The X and Y alleles were successfully detected, indicating accurate sex identification.
• Visual detection of white turbidity confirmed the presence of amplified DNA.

Conclusions:

• The loop-mediated isothermal amplification (LAMP) method provides a rapid, simple, and effective means for DNA sex determination from dental pulp.
• This technique is suitable for analyzing DNA from teeth stored for up to 25 years.
• The LAMP method holds significant potential for use in forensic identification, particularly in mass disaster scenarios.