A short interspersed nuclear element-based quantitative PCR assay for simultaneous human and dog DNA detection and...

Area of Science:

• Molecular Biology
• Forensic Science
• Genetics

Background:

• Accurate quantification of human and canine DNA is crucial for forensic casework and scientific research.
• Existing methods may not efficiently handle multispecies DNA mixtures.
• The need for a simultaneous detection assay for both species is evident.

Purpose of the Study:

• To develop a quantitative PCR (qPCR) assay for simultaneous quantification of human and dog DNA.
• To validate the assay's performance in terms of sensitivity, specificity, and precision.
• To establish a tool applicable to forensic and veterinary genetics.

Main Methods:

• Utilized the human-specific *Alu* Yb8 element and the canine-specific SINEC_Cf element as targets.
• Incorporated a universal exogenous assay with a synthetic template as an internal positive control.
• Employed quantitative PCR (qPCR) for simultaneous DNA detection and quantification.

Main Results:

• The developed qPCR assay demonstrated high sensitivity, precision, and species specificity.
• The assay achieved simultaneous quantification of human and dog DNA in a single reaction.
• A fast run time was observed, indicating efficiency for routine use.

Conclusions:

• The developed qPCR assay is a robust tool for simultaneous human and dog DNA quantification.
• This assay has direct applications in forensic investigations and veterinary genetics.
• The method provides a reliable approach for analyzing domestic pet samples.