Development and validation of I-DNA1: a 15-Loci multiplex system for identity testing

Abstract

This paper presents a system for the multiplex amplification of 15 loci, known as I-DNA1, which combines mini and midiSTR technology, with amplicon sizes ranging from 49 to 297 bp. I-DNA1 analyses all the STR loci included in the CODIS and the Interpol Standard Set of loci, nine of the ten European core loci and seven of the eight German core loci, making it suitable for use in identifying humans. Moreover, its high sensitivity and the small size of its amplicons mean that I-DNA1 is potentially highly useful for analysing highly degraded and/or very small DNA samples.

Development and validation for identity testing of I-DNADuo, a combination of I-DNA1 and a new multiplex system, I-DNA2

“…Results and discussion I-DNADuo is a combined multiplex system capable of co-amplifying 15 STR loci plus amelogenin, ten of them plus amelogenin in duplicate, in the range of 57-298 bp. It is composed of the recently developed and validated I-DNA1 multiplex system [4] and I-DNA2, a new multiplex system for the simultaneous amplification of amelogenin and 11 STR loci.…”

“…These assays were performed and amplification products were analysed by capillary electrophoresis following the methodology described in Odriozola et al [4]. Similarly, the allelic ladder for I-DNA2 was developed and its precision and accuracy were assessed via the methods used in the I-DNA1 validation by analysing 162 Caucasoid individuals (Basque Country, Spain) [4].…”

“…These primers are protected by patents P201031269 and P201031270. The regions flanking the STRs analysed were studied as described in Odriozola et al [4].…”

“…For the calculation of amplicon sizes, alleles on the STRBase web site were considered [6]. Additionally, primer pairs were chosen whose melting temperatures (58.9±0.87°C and 58.8±0.91°C, respectively) and lengths (20.4±2.3 and 21.3±2.88 bp, respectively) were similar to I-DNA1 in order to allow subsequent PCR amplification in the same thermocycling conditions for both the multiplex systems that make up I-DNADuo [4].…”

“…I-DNADuo validation is based on the validation of I-DNA1 [4] and I-DNA2 (performed following the SGWDAM guidelines) [5] to test primer set optimisation, concordance Odriozola A and Aznar JM contributed equally to this paper.…”