Abstract

Short tandem repeat (STR) loci are important polymorphic markers for paternity testing and personal identification. The AmpFLSTR SEfiler Kit commercially available is designed to amplify simultaneously eleven STR loci: D3S1358, VWA, D16S539, D2S1338, D8S1179, SE33, D19S433, TH01, FGA, D21S11, D18S51. In this study, this kit was applied to analyze the STR loci in blood/saliva samples from 255 unrelated individuals from northern Poland. Genomic DNA was isolated from the blood using a non-eznzymatic method and from the saliva using „Sherlock AX”. Multiplex PCR was performed using fluorescent primers according to the manufactures instructions – AmpFISTR®SEfiler™ users manual (Applied Biosystems). PCR products were separated by denaturing polyacrylamide capillary electrophoresis in an automated 3130 Genetic Analyzer (Applied Biosystems). Fragment sizes were determined using the internal standard GeneScan-500 LIZ. Alleles were compared with AmpFISTR SEfiler allelic ladder. The results were analyzed using GeneMapper ID v.3.2 software (Applied Biosystems).

Allele frequencies for the eleven STR systems in North Poland population and statistical parameters like PD, PIC, PE, PI, pM, heterozygosity observed (Ho) and heterozygosity expected (He) were calculated. Possible divergence from Hardy-Weinberg equilibrium was determinate by χ-square test using Utility Programs for Analysis of Genetic Linkage HWE Version 1.10 and by Exact test using Arlequin Program. The AmpFISTR®SEfiler™ has a combined power of discrimination greater than 0,99999999999999 and a cumulative power of exclusion greater than 0,99999 in the population of northern Poland.

The comparison of our data for the SE33 locus with Hungarian and northern Portugal populations revealed no significant differences (p=0,19 and p=0,61 respectively).

This paper showed that this commercial kit is useful for personal identification and paternity testing.

Adres: dr n. med. Joanna Wysocka
Katedra i Zakład Medycyny Sądowej AMG
ul. Dębowa 23, 80-204 Gdańsk
e-mail: wysocka@gumed.edu.pl