vavilovВавиловский журнал генетики и селекцииVavilov Journal of Genetics and Breeding2500-3259Institute of Cytology and Genetics of Siberian Branch of the RAS10.18699/vjgb-24-12vavilov-4060Research ArticleГЕНЕТИКА ЖИВОТНЫХANIMAL GENETICSУниверсальная панель STR-локусов для исследования полиморфизма вида Canis lupus и криминалистической идентификации собаки и волкаA universal panel of STR loci for the study of polymorphism of the species Canis lupus and forensic identification of dog and wolfhttps://orcid.org/0000-0002-1224-3275ГребенчукА. Е.HrebianchukA. E.

Минск

Minsk

iamsanya94@mail.ruhttps://orcid.org/0000-0002-8611-8215ЦыбовскийИ. С.TsybovskyI. S.

Минск

Minsk

Государственный комитет судебных экспертиз Республики БеларусьБеларусьForensic Examination Committee of the Republic of BelarusBelarusРеспубликанское унитарное предприятие «БелЮрОбеспечение»БеларусьBelJurZabespjachjenneBelarus20240203202428198107Copyright © Гребенчук А.Е., Цыбовский И.С., 20242024Гребенчук А.Е., Цыбовский И.С.Hrebianchuk A.E., Tsybovsky I.S.This work is licensed under a Creative Commons Attribution 4.0 License.https://vavilov.elpub.ru/jour/article/view/4060

Коммерческие панели микросателлитных (STR) локусов предназначены для работы с ДНК собаки домашней (Canis lupus familiaris), в связи с чем при генотипировании волка обыкновенного (Canis lupus lupus) большинство маркеров показывают существенные отклонения от равновесия Харди–Вайнберга и имеют низкий показатель информативной ценности, что осложняет их использование в судебной экспертизе. Целью настоящего исследования стал подбор STR­-маркеров, которые одинаково эффективно отображают популяционный полиморфизм волка и собаки, с последующим созданием универсальной панели для дифференциации и идентификации особей волка и собаки в криминалистике. На основе исследования полиморфизма 34 STR­-локусов сконструирована панель CPlex из 15 аутосомных локусов и двух локусов половой принадлежности, которая одинаково применима для идентификации волка и собаки. Анализ молекулярной дисперсии (AMOVA) между выборками выявил достоверные значения дифференциации (FST = 0.0828, p < 0.05), что позволяет использовать панель для дифференциации образцов волка обыкновенного и собаки домашней. Впервые в судебной экспертизе объектов животного происхождения в Республике Беларусь рассчитаны коэффициенты подразделенности популяции (θ­-value) для каждого из 15 STR­-локусов разработанной тест­-системы. Показано, что значения частот генотипов, усредненные по всем исследованным животным без учета и с учетом θ-­value, различаются на три порядка (3.39 · 10–17 и 4.71 · 10–14 соответственно). Применение коэффициентов подразделенности популяции позволит оперировать наиболее достоверными результатами экспертного идентификационного исследования. Предложенная тест-система валидирована в соответствии с протоколом Scientific Working Group on DNA Analysis Methods. Создан информационно­-статистический комплекс для автоматизации обсчета генетических данных волка обыкновенного и собаки домашней в судебной экспертизе, утверждены две методики для практикующих судебных экспертов. Методические разработки успешно применяются в экспертной практике при расследовании фактов незаконной охоты, жестокого обращения с животными и других правонарушений в Республике Беларусь.

Commercial panels of microsatellite (STR) loci are intended for DNA analysis of the domestic dog (Canis lupus familiaris) and, therefore, when genotyping the Grey wolf (Canis lupus lupus), most markers reveal significant deviations from the Hardy–Weinberg equilibrium and have a low informative value, which complicates their use in a forensic examination. The aim of this study was to select STR markers that equally effectively reflect population polymorphism in the wolf and the dog, and to create a universal panel for the identification of individuals in forensic science. Based on the study of polymorphisms of 34 STR loci, a CPlex panel of 15 autosomal loci and two sex loci was developed, which is equally suitable for identifying wolfs and dogs. Analysis of molecular variance (AMOVA) between samples revealed significant differentiation values (FST = 0.0828, p < 0.05), which allows the panel to be used for differentiating between wolf and dog samples. For the first time in the forensic examination of objects of animal origin in the Republic of Belarus, population subdivision coefficients (θ­-values) were calculated for each of the 15 STR loci of the test system being reported. It was shown that the values of the genotype frequency, when averaged over all studied animals without and with considering the θ­-value, differ by three orders of magnitude (3.39 · 10–17 and 4.71 · 10–14, respectively). The use of population subdivision coefficients will provide the researcher with the most relevant results of an expert identification study. The test system was validated in accordance with the protocol of the Scientific Working Group on DNA Analysis Methods. A computational tool was developed to automate the analysis of genetic data on the wolf and dog in the forensic examination; two guides were approved for practicing forensic experts. This methodology is being successfully used in expert practice in investigating cases of illegal hunting, animal abuse and other offenses in the Republic of Belarus.

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The authors declare that there are no conflicts of interest present.