title: Identification of a common single nucleotide polymorphism at the primer binding site of D2S1360 that causes heterozygote peak imbalance when using the Investigator HDplex Kit.
abstract: Phenomena known as null alleles and peak imbalance can occur because of mutations in the primer binding sites used for DNA typing. In these cases, an accurate statistical evaluation of DNA typing is difficult. The estimated likelihood ratio is incorrectly calculated because of the null allele and allele dropout caused by mutation-induced peak imbalance. Although a number of studies have attempted to uncover examples of these phenomena, few reports are available on the human identification kit manufactured by Qiagen. In this study, 196 Japanese individuals who were heterozygous at D2S1360 were genotyped using an Investigator HDplex Kit with optimal amounts of DNA. A peak imbalance was frequently observed at the D2S1360 locus. We performed a sequencing analysis of the area surrounding the D2S1360 repeat motif to identify the cause for peak imbalance. A point mutation (G>A transition) 136 nucleotides upstream from the D2S1360 repeat motif was discovered in a number of samples. The allele frequency of the mutation was 0.0566 in the Japanese population. Therefore, human identification or kinship testing using the Investigator HDplex Kit requires caution because of the higher frequency of single nucleotide polymorphisms at the primer binding site of D2S1360 locus in the Japanese population.
authors.affiliation: Department of Forensic Medicine, Juntendo University Graduate School of Medicine, 2-1-1 Hongo, Bunkyo, Tokyo, 113-8421, Japan. s-inoku@juntendo.ac.jp.|Criminal Scientific Investigation Laboratory, Metropolitan Police Department of Tokyo, 3-35-21, Shakujiidai, Nerima, Tokyo, 177-0045, Japan. s-inoku@juntendo.ac.jp.
authors.forename: Shota
authors.identifier: http://orcid.org/0000-0002-8051-3206
authors.initials: S
authors.lastname: Inokuchi
authors.affiliation: Criminal Scientific Investigation Laboratory, Metropolitan Police Department of Tokyo, 3-35-21, Shakujiidai, Nerima, Tokyo, 177-0045, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Yamashita
authors.affiliation: Criminal Scientific Investigation Laboratory, Metropolitan Police Department of Tokyo, 3-35-21, Shakujiidai, Nerima, Tokyo, 177-0045, Japan.
authors.forename: Kazuma
authors.initials: K
authors.lastname: Nishimura
authors.affiliation: Department of Forensic Medicine, Juntendo University Graduate School of Medicine, 2-1-1 Hongo, Bunkyo, Tokyo, 113-8421, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Nakanishi
authors.affiliation: Department of Forensic Medicine, Juntendo University Graduate School of Medicine, 2-1-1 Hongo, Bunkyo, Tokyo, 113-8421, Japan.
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Saito
mesh_terms: D044466:Asian People / Q000235:genetics; D016172:DNA Fingerprinting / Q000295:instrumentation*; D017931:DNA Primers; D005787:Gene Frequency; D005838:Genotype; D006579:Heterozygote; D006801:Humans; D007564:Japan; D017354:Point Mutation; D020641:Polymorphism, Single Nucleotide; D060888:Real-Time Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
chemical_list: D017931:DNA Primers
keywords: D2S1360; Investigator HDplex; Peak imbalance; Primer binding site mutation; Short tandem repeat