title: The application of an automated allele concordance analysis system (CompareCalls) 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 (CompareCalls, 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 SurelockID 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 SurelockID 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.
authors.affiliation: Myriad Genetic Laboratories, 320 Wakara Way, Salt Lake City, Utah 84108, USA.
authors.forename: John H
authors.initials: JH
authors.lastname: Ryan
authors.forename: Jeffrey K
authors.initials: JK
authors.lastname: Barrus
authors.forename: Bruce
authors.initials: B
authors.lastname: Budowle
authors.forename: Cynthia M
authors.initials: CM
authors.lastname: Shannon
authors.forename: Victor W
authors.initials: VW
authors.lastname: Thompson
authors.forename: Brian E
authors.initials: BE
authors.lastname: Ward
mesh_terms: D000483:Alleles; D001331:Automation; D016172:DNA Fingerprinting / Q000379:methods / Q000592:standards*; D004586:Electrophoresis; D005819:Genetic Markers; D005838:Genotype; D006801:Humans; D011786:Quality Control; D015203:Reproducibility of Results; D012984:Software; D020080:Tandem Repeat Sequences
chemical_list: D005819:Genetic Markers