Abstract:
Commercial panels of microsatellite (STR) loci are focused on the use of DNA of the domestic dog (Canis lupus familiaris) and are often inapplicable for genotyping the DNA of the gray wolf (Canis lupus lupus). We propose a CPlex test system, including one hexa- and 12 tetranucleotide autosomal STR loci, as well as two sex loci, that is equally efficient in DNA identification of biological samples of the wolf and the dog. Analysis of molecular variance between samples revealed significant differentiation values (FST = 0.0784, p < 0.001), which allows to use the panel to differentiate wolf and dog samples. Population subdivision coefficients (θ-values) were calculated for each of the 13 STR loci of the developed test system. It was shown that the values of the genotype frequency for dogs and wolves, without and with considering the θ-value, differ by three orders of magnitude (for dogs 8.9 × 10-16 and 2.1 × 10-14 and for wolves 1.9 × 10-15 and 4.5 × 10-14, respectively). The use of population subdivision coefficients will allow to identify the most reliable results of an expert identification study and the power of exclusion provided by the STR loci of the CPlex test system makes it possible to achieve a reliable level of evidence in forensic DNA analysis of both wolves and dogs. The test system has been validated for use in forensic identification of the dog and wolf based on biological traces found at crime scenes, as well as for individual identification and establishing biological relationship of animals of these species.
摘要:
微卫星(STR)基因座的商业面板专注于使用家犬(犬类狼疮)的DNA,并且通常不适用于对灰狼(犬类狼疮)的DNA进行基因分型。我们提出了一个CPlex测试系统,包括一个六核苷酸和12个四核苷酸常染色体STR基因座,以及两个性别位点,这在狼和狗的生物样本的DNA鉴定中同样有效。样品之间的分子方差分析揭示了显著的差异值(FST=0.0784,p<0.001),它允许使用面板来区分狼和狗的样本。为开发的测试系统的13个STR基因座中的每一个计算了种群细分系数(θ值)。结果表明,狗和狼的基因型频率值,在不考虑θ值的情况下,相差三个数量级(狗分别为8.9×10-16和2.1×10-14,狼分别为1.9×10-15和4.5×10-14)。人口细分系数的使用将允许确定专家鉴定研究的最可靠结果,并且CPlex测试系统的STR基因座提供的排除能力使在法医DNA分析中获得可靠的证据成为可能。狼和狗。该测试系统已通过验证,可根据犯罪现场发现的生物痕迹,对狗和狼进行法医鉴定,以及这些物种动物的个体识别和建立生物关系。