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01-05-2021 | Original Article

A novel forensic panel of 186-plex SNPs and 123-plex STR loci based on massively parallel sequencing

Authors: Xinyao Miao, Yuesheng Shen, Xiaojuan Gong, Huiyun Yu, Bowen Li, Liao Chang, Yinan Wang, Jingna Fan, Zuhuan Liang, Bowen Tan, Shengbin Li, Bao Zhang

Published in: International Journal of Legal Medicine | Issue 3/2021

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^TM, 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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Title: A novel forensic panel of 186-plex SNPs and 123-plex STR loci based on massively parallel sequencing
Authors: Xinyao Miao
Yuesheng Shen
Xiaojuan Gong
Huiyun Yu
Bowen Li
Liao Chang
Yinan Wang
Jingna Fan
Zuhuan Liang
Bowen Tan
Shengbin Li
Bao Zhang
Publication date: 01-05-2021
Publisher: Springer Berlin Heidelberg
Published in: International Journal of Legal Medicine / Issue 3/2021
Print ISSN: 0937-9827
Electronic ISSN: 1437-1596
DOI: https://doi.org/10.1007/s00414-020-02403-z

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A novel forensic panel of 186-plex SNPs and 123-plex STR loci based on massively parallel sequencing

Abstract

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Abstract

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