AI Article Synopsis

  • The study introduces a new forensic panel utilizing the BGISEQ-500 platform to analyze 186 single-nucleotide polymorphisms (SNPs) and 123 short tandem repeats (STRs) for forensic applications, addressing a gap in forensic capabilities with this sequencing technology.
  • Researchers confirmed the panel's effectiveness by evaluating critical factors such as accuracy, sensitivity, and repeatability, achieving 100% accurate genotypes with a high detection rate of over 98% from low amounts of input DNA.
  • The results indicate that the panel is versatile and reliable for various forensic needs, including paternity testing and analyzing degraded samples, making it a valuable advancement in forensic science methodologies.

Article Abstract

The MiSeq® FGX Forensic system and the HID-Ion AmpliSeq Panel were previously developed for massively parallel sequencing (MPS) for forensic casework. Among the three major sequencing platforms, BGISEQ-500, which is based on multiple PCRs, is still lacking in forensics. Here, a novel forensic panel was constructed to detect 186 single-nucleotide polymorphisms (SNPs) and 123 short tandem repeats (STRs) with MPS technology on the BGISEQ-500™ platform. First, the library preparation, sequencing process, and data analysis were performed, focusing on the average depth of coverage and heterozygote balance. We calculated the allelic frequencies and forensic parameters of STR and SNP loci in 73 unrelated Chinese Han individuals. In addition, performance was evaluated with accuracy, uniformity, sensitivity, PCR inhibitor, repeatability and reproducibility, mixtures, degraded samples, case-type samples, and pedigree analyses. The results showed that 100% accurate and concordant genotypes can be obtained, and the loci with an abundance in the interquartile range accounted for 92.90% of the total, suggesting reliable uniformity in this panel. We obtained a locus detection rate that was higher than 98.78% from 78 pg of input DNA, and the optimal amount was 1.25-10 ng. The maximum concentrations of hematin and humic acid were 200 and 100 μM, respectively (the ratios of detected loci were 96.52% and 92.41%), in this panel. As a mixture, compared with those of SNPs, minor-contributor alleles of STRs could be detected at higher levels. For the degraded sample, the ratio of detected loci was 98.41%, and most profiles from case-type samples were not significantly different in abundance in our studies. As a whole, this panel showed high-performance, reliable, robust, repeatable, and reproducible results, which are sufficient for paternity testing, individual identification, and use for potentially degraded samples in forensic science.

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http://dx.doi.org/10.1007/s00414-020-02403-zDOI Listing

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