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Development and validation of a multiplex insertion/deletion marker panel, SifaInDel 45plex system.

Ruiyang Tao1, Jingyi Zhang2, Xiang Sheng3

  • 1Institute of Forensic Medicine, West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, Chengdu, 610041, PR China; Shanghai Key Laboratory of Forensic Medicine, Shanghai Forensic Service Platform, Academy of Forensic Sciences, Ministry of Justice, Shanghai, 200063, PR China.

Forensic Science International. Genetics
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Summary

The SifaInDel 45plex system efficiently amplifies 45 insertion and deletion polymorphisms (InDels) for forensic identification. This robust multiplex panel provides high discrimination power for human identification and kinship analysis.

Keywords:
Capillary electrophoresis (CE)DNA genotypingForensic scienceInsertion and deletion polymorphisms (InDels)

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Area of Science:

  • Forensic Genetics
  • Molecular Biology
  • Human Identification

Background:

  • Insertion and deletion polymorphisms (InDels) offer advantages over short tandem repeats (STRs) and single nucleotide polymorphisms (SNPs) in forensic genetics.
  • A need exists for multiplex systems capable of simultaneously analyzing a large number of InDel markers for enhanced human identification.

Purpose of the Study:

  • To design and validate the SifaInDel 45plex system for simultaneous amplification and detection of 45 InDel markers.
  • To evaluate the performance of the SifaInDel 45plex panel in terms of sensitivity, specificity, and population genetics for forensic applications.

Main Methods:

  • Development of a single polymerase chain reaction (PCR) procedure to amplify 27 autosomal InDels (A-InDels), 16 X chromosome InDels (X-InDels), and 2 Y chromosome InDels (Y-InDels).
  • Detection of amplified products using capillary electrophoresis (CE).
  • Validation studies including optimization of PCR conditions, repeatability, reproducibility, concordance, accuracy, sensitivity, stability, species specificity, and population genetics analysis.

Main Results:

  • The SifaInDel 45plex panel successfully generated full DNA profiles from as little as 62.5 pg of input DNA and from challenging casework samples.
  • The system demonstrated tolerance to common inhibitors (hematin, nigrosine, urea).
  • High combined power of discrimination (CPD > 0.999999 for A-InDels) and exclusion (CPED = 0.955118, CPET = 0.997754) were achieved. X- and Y-InDels also showed high discriminatory power and utility for gender determination.

Conclusions:

  • The SifaInDel 45plex panel is a reliable and robust multiplex system for forensic human identification and complex kinship analysis.
  • The panel's ability to analyze a large number of autosomal, X, and Y chromosome InDels on a CE platform makes it a valuable complementary tool in forensic genetics.