Related Experiment Video
Updated: Jan 14, 2026
08:44
A High-Throughput In Situ Method for Estimation of Hepatocyte Nuclear Ploidy in Mice
Published on: April 19, 2020
10.4K
Accurate Detection and Quantification of Human Nuclear DNA
Toshifumi Nakagawa1, Masanori Doi2, Kosuke Nishi3
1Forensic Science Laboratory, Ehime Prefectural Police Headquarters, 2-2 Minamihoribatacho, Matsuyama, Ehime 790-8573, Japan; Faculty of Agriculture, Ehime University, 3-5-7 Tarumi, Matsuyama, Ehime 790-8566, Japan.
Legal Medicine (Tokyo, Japan)
|October 24, 2025
Summary
A new human nuclear DNA (nDNA) assay accurately identifies human origin and quantifies nDNA for forensic DNA profiling. This HAR1 assay is specific to humans and works even with limited or mixed DNA samples.
Area of Science:
- Forensic Science
- Molecular Biology
- Genetics
Background:
- Accurate human DNA profiling is crucial in forensic science.
- Existing methods can be challenged by limited sample quantity/quality or non-human DNA contamination.
- Optimizing DNA input for PCR is essential for reliable results.
Purpose of the Study:
- To develop a novel assay for simultaneous human DNA origin confirmation and quantification.
- To validate the specificity and sensitivity of the assay across various species and sample types.
- To assess the assay's performance in the presence of non-human DNA.
Main Methods:
- Development of a TaqMan probe-based real-time PCR assay targeting the human accelerated region 1 (HAR1).
- Testing specificity against 23 vertebrate species.
- Quantification of human nDNA from single somatic cell equivalents.
- Evaluation of assay performance with mixed human and non-human DNA samples.
Main Results:
- The HAR1 assay demonstrated exclusive specificity to human DNA, with no cross-reactivity in 23 tested vertebrate species.
- Accurate human nDNA detection was achieved from as little as a single somatic cell.
- Precise quantification of human nDNA was obtained down to 33 pg/μL, meeting commercial kit recommendations.
- Non-human DNA did not interfere with the accuracy of human nDNA quantification.
Conclusions:
- The HAR1 assay is a highly specific and sensitive tool for confirming human origin in forensic samples.
- It enables accurate quantification of human nDNA, optimizing input for DNA profiling.
- This assay enhances the efficiency and reliability of forensic DNA analysis, especially with challenging samples.
