title: Improving DNA data exchange: validation studies on a single 6 dye STR kit with 24 loci.
abstract: The idea of developing a new multiplex STR amplification system was conceived in 2011 as an effective way to implement the new European standard set (ESS) of 12 STR markers adopted by The Council of the European Union in 2009 while maintaining an effective compatibility and information exchange with the historical DNA profiles contained in the Spanish national DNA database (around 200,000 DNA profiles) mainly based on the 13 CODIS core STR loci plus D19S433 and D2S1338 markers. With this goal in mind we proposed to test and validate a single STR amplification system for simultaneous analysis of 21 STR markers covering both CODIS and ESS core STR loci plus three additional markers (D19S433, D2S1338, and SE33) also contained in commonly used STR kits and national DNA databases. In 2012, we started the first beta-testing with a 6-dye STR kit prototype containing 24 loci (now known as the GlobalFiler™ PCR Amplification Kit) developed by Life Technologies in response to the CODIS Core Loci Working Group's recommendation to expand the CODIS Core Loci. This prototype included our proposal of 21 autosomal STR markers and two Y-chromosome markers (DYS391 and Y-indel) and maximizes concordance with established databases and previously analyzed samples by maintaining primer sequences of previous Identifiler(®)/NGM SElect™ kits for the 21 STR markers except for TPOX. This paper describes the validation studies conducted with the first commercial available 6-dye STR kit for casework using a 3500 genetic analyzer for fragment detection that included the analysis of the following parameters and aspects: analytical threshold, sensitivity & stochastic threshold, heterozygous balance, stutter threshold, precision and accuracy, repeatability and reproducibility, genotype concordance, DNA mixtures, species specificity, and stability studies with case type samples. The studies demonstrated that the GlobalFiler™ system provided equivalent overall performance to previous forensic STR PCR kits, but with enhanced discrimination power for a better match efficiency that would reduce the chance of adventitious matches during DNA data exchange among national DNA databases.
authors.affiliation: National Institute of Toxicology and Forensic Sciences, Madrid Department, Biology Service, José Echegaray 4, Las Rozas 28232, Madrid, Spain.
authors.forename: Pablo
authors.initials: P
authors.lastname: Martín
authors.affiliation: National Institute of Toxicology and Forensic Sciences, Madrid Department, Biology Service, José Echegaray 4, Las Rozas 28232, Madrid, Spain.
authors.forename: Lourdes Fernández
authors.initials: LF
authors.lastname: de Simón
authors.affiliation: National Institute of Toxicology and Forensic Sciences, Madrid Department, Biology Service, José Echegaray 4, Las Rozas 28232, Madrid, Spain.
authors.forename: Gracia
authors.initials: G
authors.lastname: Luque
authors.affiliation: National Institute of Toxicology and Forensic Sciences, Madrid Department, Biology Service, José Echegaray 4, Las Rozas 28232, Madrid, Spain.
authors.forename: María José
authors.initials: MJ
authors.lastname: Farfán
authors.affiliation: National Institute of Toxicology and Forensic Sciences, Madrid Department, Biology Service, José Echegaray 4, Las Rozas 28232, Madrid, Spain. Electronic address: antonio.alonsoalonso@justicia.es.
authors.forename: Antonio
authors.initials: A
authors.lastname: Alonso
mesh_terms: D041322:Chromosomes, Human, Y / Q000235:genetics; D016172:DNA Fingerprinting / Q000295:instrumentation*; D030561:Databases, Nucleic Acid; D005260:Female; D005819:Genetic Markers; D006801:Humans; D033181:Information Dissemination; D008297:Male; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction / Q000295:instrumentation*
chemical_list: D005819:Genetic Markers
keywords: 6-Channel detection systems; Autosomal STR markers; DNA data exchange; Forensic validation studies; GlobalFiler™ system