Evaluation of the MiSeq FGx system for use in forensic casework
Capillary electrophoresis (CE) is widely used in forensic genetics to study short tandem repeats (STRs). Recently, next-generation sequencing (NGS) platforms have facilitated the development of new strategies for forensic DNA typing. Several studies have shown that NGS successfully analyzes challenging samples. However, because NGS is complicated and time-consuming, it remains unclear whether NGS platforms offer significant advantages over CE for all forensic cases. Here, the MiSeq FGx system was used to test some cases that had previously been analyzed using CE. These cases included paternity test cases in which some samples exhibited locus inconsistencies; samples with off-ladder (OL) alleles; samples with triallelic patterns; and samples with amelogenin test abnormalities. The results generated by MiSeq FGx were compared to those previously generated by CE. The MiSeq FGx and CE results were consistent with the exception of three samples, where inconsistencies were observed at the Penta D locus. For all three incongruent samples, the MiSeq FGx results were correct. Sequence analysis indicated that, in two cases, mismatches were due to undetected alleles rather than mutations. In two additional cases, mutation sources were identified, and in a fifth case, mutation step size was reconsidered. MiSeq FGx was used to identify OL alleles and samples with amelogenin test abnormalities. For cases where verification was required via CE analysis, the simultaneous NGS amplification of several types of multiple genetic markers improved testing efficiency. In addition, we identified additional sequence variants at autosomal, Y chromosomal, and X chromosomal STR loci in the Han Chinese population from northern China. Our results will be useful for future forensic analyses of STR genotypes in Chinese populations. It is likely that NGS would be more widely used in forensic genetics if costs and procedure complexity were reduced.
KeywordsIllumina MiSeq FGx™ system Next-generation sequencing Sequence variation Forensic genetics STR Forensic casework
The authors wish to thank Xu Liu and Jing-jing Zhang. This work was supported by the Beijing Center for Physical and Chemical Analysis, Beijing, People’s Republic of China.
This work was supported in part by a grant from the Millions of Talent Projects, China. The sponsor had no involvement in the development of the paper or decisions related to the paper.
Compliance with ethical standards
This study was performed in accordance with the ethical standards as laid down in the 1964 Declaration of Helsinki and its later amendments.
Conflict of interest
The authors declare that they have no conflict of interest.
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