Internal Validation of Global Filer PCR Amplification Kit in DNA Section of Punjab Forensic Science Agency, Lahore

Abstract

Background: Global Filer PCR Amplification Kit is a 6-dye amplification assay, which amplifies 24 genetic loci simultaneously in a single reaction. The amplification system encompasses both Combined DNA Index System loci and European standard set loci along with a highly polymorphic short tandem repeat (STR) locus SE33. The addition of a Y-STR marker and an insertion deletion marker of Y chromosome has provided additional aid in the gender identification of Amelogenin-Y null males. Global Filer PCR Amplification Kit has provided understandably more discrimination power compared with Identifiler Plus PCR Amplification Kit currently being used in Punjab Forensic Science Agency (PFSA). Methodology: In this study, Global Filer PCR Amplification Kit was internally validated in PFSA following the Scientific Working Group on DNA Analysis Methods laid principles to establish quality assurance parameters and interpretation guidelines. Known nonprobative DNA samples were utilized in this study to determine the sensitivity, establishment of proper thresholds for analysis, mixture analysis to determine detectable mixture ratios, contamination assessment, and reproducibility for the new method. Conclusion: The results of this internal validation strongly suggest the use of Applied Biosystems Global Filer PCR Amplification Kit in forensic casework analysis.