VALIDATION OF STR-LOCUS FRAGMENT ANALYSIS METHOD BY USING A SET OF REAGENTS AmpFISTR® IDENTIFIER ® PLUS FOR CARRYING OUT THE AMPLIFICATION REACTION

Abstract

The purpose of the article is to clarify the results of in-vitro validation method of the fragmentation analysis of the STR-locus on the AB 3130 and 3100-Avant genetic analyzers, using the AmpFlSTR® Identifiler ® Plus reagent kit for the  amplification reaction, and to provide specific recommendations. During the study several parameters have been checked (accuracy and reliability of the method, its sensitivity, optimal DNA concentration, heterozygous balance, stator-allele ratio in the DNA profile, additional peaks (+ A, −A) and atypical alleles (artifacts) were tested within the study, the effect of the inhibitors on the amplification reaction, the quality of the amplification reaction during the degradation DNA study, the reliability of the results according to NIST DNA standards, the DNA mixture at different ratios of DNA concentration, analytical and stochastic thresholds). The accuracy of the method of fragment analysis of STR-loci using the set of Identifiler® Plus reagents and the high level of reliability of the results of the study, its complete suitability and efficacy for establishing the genetic traits of a person (corpse), as well as biological traces, were confirmed including the  confirmation of the method in the presence of inhibitors (with the exception of degraded DNA studies). The methodological basis of the study includes laws and categories of the theory of cognition, particularly the provisions of materialistic dialectics, which contributed to the awareness of the object, subject, purpose and objectives of the study in the context of the interdependence of achievements and needs of practice. At the same time, formal logic methods (analysis,
synthesis, deduction, induction, analogy, abstraction) made it possible to understand in more detail the content of the investigated issues, and the essence of the studied categories and phenomena was clarified by the system-structural method. The simulation method was used to outline the flow of in vitro laboratory validation of the fragment analysis of STR loci method on genetic analyzer AB 3130 and 3100-Avant using the AmpFlSTR® Identifiler® Plus reagent kit for carrying out the amplification reaction.
Keywords: validation; deoxyribonucleic acid; short tandem repetition; polymerase chain reaction; fragment capillary electrophoresis.