Abstract
Short tandem repeat (STR) profiling has been routinely used in kinship testing since the introduction of commercial kits in the mid-1990s. While 15 to 23 STR loci normally give definitive results in simple kinship testing, additional loci are sometimes required to resolve complex cases. The SureID 23comp Human Identification Kit, recently released by Health Gene Technologies (China), multiplexes amelogenin and 22 autosomal STRs, 17 of which are non-CODIS STRs. This enables the profiling of 38–40 loci when used in conjunction with widely used commercial kits. In this study, the kit was evaluated for kinship applications as a supplementary STR kit following the minimum criteria for validation recommended by the European Network of Forensic Science Institutes (ENFSI) and the Scientific Working Group on DNA Analysis Methods (SWGDAM) using 500 samples. Performance was comparable with other commercial kits demonstrating: repeatability and reproducibility; precision (maximum s.d. 0.1048 nt); accuracy, all alleles were within ±0.41 nt compared to the actual sizes; heterozygous peak balances at all loci >68%; stutter ratios ranged from 3.8% to 16.15%; full profiles were generated with 125 pg DNA (95.12% of alleles at 62 pg),; and we found 100% concordance over 5 common STRs with the GlobalFiler kit.
Highlights
The extended number of Short tandem repeat (STR) markers required for the Combined DNA Index System (CODIS) and for the European Standard Set (ESS)[1,2], has led to the development of GlobalFiler PCR Amplification Kit, VeriFiler Plus PCR Amplification Kit (Applied Biosystems (AB), USA), PowerPlex Fusion 6 C System (Promega Corporation, USA) and Investigator 24plex (Qiagen, Germany)
The first step in the validation was a confirmation of the identity of the D5 locus included in the multiplex by testing the 9947 A control DNA provided in the kit as a positive control (Note 1)
Full profiles were generated from the 125 pg samples when using the 10 μl volume (28 and 30 cycles), while 25 μl volume was able to generate a full profile with 30 PCR cycles only
Summary
The extended number of STR markers required for the Combined DNA Index System (CODIS) and for the European Standard Set (ESS)[1,2], has led to the development of GlobalFiler PCR Amplification Kit, VeriFiler Plus PCR Amplification Kit (Applied Biosystems (AB), USA), PowerPlex Fusion 6 C System (Promega Corporation, USA) and Investigator 24plex (Qiagen, Germany). A set of 25 supplementary STRs was suggested by the National Institute of Standards and Technology (NIST) to increase the certainty in kinship testing[8]; no multiplex combing these STRs is commercially available. These can be utilised to increase the power of kinship testing[15,16]. The systems are expensive to establish and are not yet commonly used in many laboratories
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