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International Journal of Legal Medicine (2023) 137:1-12
https://doi.org/l 0.1007/s00414-022-02907-w
ORIGINAL ARTICLE
Development of a multiplex panel with 31 multi-allelic InDels
for forensic DNA typing
Yining Yao - Kuan SunZ3 - Qinrui Yang' - Zhihan Zhou - Jinglei Qian' - Zhimin Li1 - Chengchen Shao -
Xiaoqin Qian' - Qiqun Tang4 - Jianhui Xie1
Received: 15 March 2022 / Accepted: 20 October 2022 / Published online: 3 November 2022
© The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2022
Abstract
Insertion/Deletion (InDel) polymorphic genetic markers are abundant in human genomes. Diallelic InDel markers have
been widely studied for forensic purposes, yet the low polymorphic information content limits their application and cur-
rent InDel panels remain to be improved. In this study, multi-allelic InDels located out of low complexity sequence regions
were selected in the datasets from East Asian populations, and a multiplex amplification system containing 31 multi-allelic
InDel markers and the Amelogenin marker (FA-HID32plex) was constructed and optimized. The preliminary study on
sensitivity, species specificity, inhibitor tolerance, mixture resolution, and the detection of degraded samples demonstrates
that the FA-HID32plex is highly sensitive, specific, and robust for traces and degraded samples. The combined power of
discrimination (CPD) of 31 multi-allelic InDel markers was 0.999 999 999 999 999 999 85, and the cumulative probability
of exclusion (CPE) was 0.999 920 in a Chinese Han population, which indicates a high discrimination power. Altogether,
the FA-HID32plex panel could provide reliable supplements or stand-alone information in individual identification and
paternity testing, especially for challenging samples.
Keywords Forensic Genetics - Multi-allelic InDel - Multiplex amplification - Capillary electrophoresis - Degraded samples
Introduction
Insertion/Deletion (InDel) polymorphic genetic markers
are length polymorphisms with the insertion or deletion of
short DNA sequences. Evidence has shown that InDels are
Yining Yao and Kuan Sun contributed equally to this work.
Jianhui Xie
jhxie@fudan.edu.cn
Department of Forensic Medicine, School of Basic
Medical Sciences, Fudan University, 138 Yixueyuan Road,
Shanghai 200032, China
2 Department of Fetal Medicine and Prenatal Diagnosis
Center, Shanghai First Maternity and Infant Hospital, Tongji
University School of Medicine, 2699 West Gaoke Rd,
201204 Shanghai, China
3 Shanghai Key Laboratory of Maternal Fetal Medicine,
Shanghai First Maternity and Infant Hospital, School
of Medicine, Tongji University, Shanghai 200092, China
4 Department of Biochemistry and Molecular Biology,
School of Basic Medical Sciences, Fudan University,
Shanghai 200032, China
abundant in human genomes [1, 2]. Generally, InDel mark-
ers could be classified into five categories according to their
length and sequence types, which are insertions and dele-
tions of single-base pairs, monomeric base pair expansions,
multi-base pair expansions of 2-15 bp repeat units, trans-
poson insertions, and InDels containing apparently random
DNA sequences [3]. The plausible formation of InDels could
be explained by polymerase slippage and a series of mecha-
nisms for cellular DNA repair against secondary structure,
double-strand DNA break, defective mismatch, and unequal
meiotic recombination [4-7].
Studies have been carried out on the screening and inves-
tigation of InDel markers for forensic purposes [8-11].
Advantages have been gained on the detection of degraded
biological samples using InDels since InDel markers present
a shorter amplicon than short tandem repeat (STR) markers
[8, 12]. Stable inheritance could be ensured by the relatively
lower mutation rate in InDel polymorphisms [13]. In addi-
tion, the detection of InDels appears easy and economi-
cal with capillary electrophoresis (CE) analysis due to the
length polymorphic trait of InDel markers. Altogether, InDel
1 Springer](ViewImageLocal?handle=hein.journals/injlegame137&div=1&collection=&method=preview&ext=.png&size=3)
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