Related Experiment Video
Updated: Oct 21, 2025
05:53
Candidate Gene Testing in Clinical Cohort Studies with Multiplexed Genotyping and Mass Spectrometry
Published on: June 21, 2018
10.3K
Development and Validation of a Forensic Multiplex System With 38 X-InDel Loci
Ling Chen1, Xiyong Pan2, Yuan Wang1
1School of Forensic Medicine, Southern Medical University, Guangzhou, China.
Frontiers in Genetics
|September 6, 2021
Summary
A new multiplex kit, AGCU X-InDel 38, efficiently amplifies 38 X-InDel markers for forensic DNA analysis. It demonstrates high sensitivity, inhibitor tolerance, and robust performance in challenging kinship cases.
Area of Science:
- Forensic Science
- Genetics
- Molecular Biology
Background:
- Forensic DNA analysis relies on polymorphic markers for individual identification and kinship testing.
- Existing multiplex systems face challenges with degraded DNA, low template amounts, and common inhibitors.
- X-InDel markers offer advantages for specific forensic applications, including sex determination and kinship analysis.
Purpose of the Study:
- To develop and validate a novel multiplex system, the AGCU X-InDel 38 kit, for amplifying 38 X-InDel markers and amelogenin.
- To assess the kit's suitability and efficiency for forensic applications, including sensitivity, specificity, and reproducibility.
- To evaluate the kit's performance with challenging samples, such as degraded and trace DNA, and assess its discriminatory power in a specific population.
Main Methods:
- Design and validation of the AGCU X-InDel 38 kit for simultaneous amplification of 38 X-InDel markers and amelogenin using Polymerase Chain Reaction (PCR).
- Conducting a series of validation experiments covering sensitivity (down to 62.5 pg DNA), inhibitor tolerance (hematin, hemoglobin), species specificity, reproducibility, and stability.
- Analyzing case samples, including degraded and trace DNA, and evaluating allele frequencies and forensic parameters (e.g., Probability of Discrimination) in the Guangdong Han population.
Main Results:
- The AGCU X-InDel 38 kit achieved full DNA profiles with as little as 62.5 pg of template DNA.
- The kit demonstrated good tolerance to common inhibitors like hematin (≤25 μmol/L) and hemoglobin (≤50 μmol/L).
- High discriminatory power was observed, with combined Probability of Discrimination (PD) values exceeding 0.999999999 for both males and females, and combined Mean Exclusion Chances (MEC) values also indicating robust performance.
Conclusions:
- The AGCU X-InDel 38 kit is a sensitive, precise, reproducible, and highly informative tool for forensic DNA analysis.
- The kit exhibits excellent performance with degraded and trace DNA samples, outperforming in challenging casework.
- This novel multiplex system serves as a valuable complementary tool for resolving complex and challenging kinship cases.
