The Application of an Automated Allele Concordance Analysis System (Compare CallsSM) to Ensure the Accuracy of Single-Source STR DNA Profiles

Abstract

A powerful method for validating a scientific result is to confirm specific results utilizing independent methodologies and processing pathways. Thus, we have designed, developed and validated an automated allele concordance analysis system (CompareCallSM, patent pending) that performs comparisons between two independent DNA analysis platforms to ensure the highest accuracy for allele calls. Application of this system in a quality assurance role has shown the potential to eliminate greater than 90% of the STR analysis required of a DNA data analyst. While this system is broadly applicable for use with any two independent STR analysis programs, either prior to or following human data review, we are presenting its application to data generated with the ABI Prism™ Genotyper™ software system versus data generated with the SurelockIDSM system. With the automated allele concordance analysis system, the GeneScan™ DNA fragment data generated from an ABI 377™ gel image are analyzed in two independent pathways. In one analysis pathway, the GeneScan™ data are imported into Genotyper™ software where STR labels are assigned to the fragment data based upon the criteria of the Kazam 20% macro. The “Kazam” macro provided with the Genotyper™ program works by labeling all peaks in a category (or locus) and then filtering (or removing) the labels from peaks, such as those in stutter positions, that meet predefined criteria. In the second pathway, the GeneScan™ data are imported into the SurelockIDSM analysis platform where STR labels and error messages are assigned to the fragment data based upon hard-coded allele calling criteria and quality parameters. The resulting STR allele calls for each analysis platform are then compared, utilizing the automated allele concordance analysis system. Any differences in the STR allele calls between the two systems are flagged in a discordance report for further review by a qualified DNA data analyst. The automated allele concordance analysis system guides the DNA data analyst to the discordant data generated by either analysis platform. Additionally, the analyst is also directed to data that are of less than pristine quality which may have an increased potential for errors in interpretation by either analysis platform or by a human DNA data analyst. Implementation of an automated allele concordance analysis system will yield high-quality data for CODIS and free the human DNA data analyst to perform other critical duties within the laboratory.

Forensic Science

“…In a comparison of alleles calls from 2048 STR profiles between manual review with Genotyper and automated review with TrueAllele, only one significant difference was observed when the analyst using Genotyper interpreted a spike as a DNA peak at D8S1179, but TrueAllele correctly designated it as a spike (85). Validation studies with 290 676 STR markers found CompareCalls SM software, an automated allele concordance analysis system from Myriad Genetics, to be at least as accurate as 100% human technical review of STR profiles (86).…”