Validation of the AmpF lSTR ® Identifiler ® Direct PCR Amplification kit in a laboratory accredited according to the ISO17025 standard

Abstract

A total of 249 FTA ® card samples were investigated with the Identifiler ® Direct kit. PCR amplification was performed according to the manufacturer's protocol with modifications in the reaction volume (10–25 μL), the number of PCR cycles (25–27) and the number of FTA-card punches (1–2). Full concordance was observed between the results obtained with the Identifiler ® Direct kit and the washed punches typed with the Identifiler ® kit. The success rate with one 1.2 mm punch in a 15 μL PCR reaction volume and 25 PCR-cycles was 93.3%. After retyping of the failed samples, the combined success rate was 98.7%. Overall, the quality of the results was reduced when the number of punches was increased or the PCR reaction volume was decreased. The Identifiler ® Direct master mix may be used in combination with other primer sets for direct amplification of other loci, e.g. the 49 SNPs using the SNP forID multiplex assay.