Розроблена діагностична система скринінгу ендогенного ретровірусу свиней підтипу С (PERV-C) за допомогою мультиплексної ПЛР-SSP для виявлення особин із зниженим ризиком біологічної небезпеки при їх застосуванні для цілей ксенотрансплантації.На зразках ДНК, отриманих від тварин свиней порід в’єтнамський мейшан та велика біла, визначена чутливість та специфічність тест-системи PERV-C – α-Actin. Встановлено, що гранично допустимою концентрацію геномної ДНК для виявлення фрагменту ретровірусу свиней в ПЛР з наступним розділенням продуктів ампліфікації шляхом горизонтального електрофорезу в агарозному гелі є 15,2 пг/мкл, а мінімальна кількість ПЛР-продукту для можливості його візуалізації склала 5 × 103 копій.

The article presents the analysis of the PERV retrovirus subtypes A and C frequency in populations of Ukrainian and foreign breed pigs. Different frequencies of the PERV A/C genome presence in animals of the studied breeds were established. The largest relative number was observed in the group of wild pigs (86%), the smallest was in the groups of Poltava meat and Piétrain breeds. Animals free of both virus subtypes were found in all study groups. The article considers the hypothesis of an increase in the frequency of PERV retrovirus in the pigs’ genome during domestication. Its integration caused a gene mutation responsible for fat deposition which led to increased fat amount in carcasses and could be picked up by selection in the process of creating breeds. However, there is no obvious link between the spread of the virus in modern breeds in different areas of productivity. Also, there is no association between carcass fat amount and the presence of PERV in the genome. It is established that the information on the PERV A/C distribution in pig breeds hold in Ukraine is useful in terms of the possibility of using each of them for xenotransplantation. Also, this information can be used to justify the selection of founding breeds in order to create lines of pigs free from the endogenous retrovirus genome.

An important problem in transplant medicine is the lack of organs and tissues for human transplantation. One of the promising areas of its solution today is the use of organs and tissues of animals — xenotransplantation. Pigs for medical and biological needs must be stress-resistant and adapted to the conditions of keeping and feeding within the vivarium. Stress syndrome in pigs is caused by a point mutation that replaces 1843CT in exon 17 of the ryanodine RYR1 receptor gene in a homozygous state. The aim of the study was to examine the genetic structure of individual pig populations at the locus of the RYR1 gene responsible for hypersensitivity. DNA testing of 102 pigs of Poltava meat, Myrhorod, Great White, Landrace and Pietren breeds was performed. The studies were performed on DNA samples obtained from pig blood. DNA was isolated using Chelex 100 reagent. Studies of the RYR1 gene fragment were performed by restriction fragment length polymorphism (RFPR) by polymerase chain reaction (PCR). Two alleles of the ryanodine RYR1 receptor gene (n and N) and three genotypes (NN, Nn, nn) were identified in the studied animals. A molecular study of pigs of different breeds revealed a polymorphism of the RYR1 gene. The number of animals carrying the recessive allele n of the ryanodine receptor gene RYR1, which is responsible for the sensitivity of pigs to stress factors, varied considerably: from its complete absence in the Vietnamese Pot-bellied, Ukrainian meat, Large White and Ukrainian Spotted Steppe breeds up to 100% of animals of homozygous RYR1nn genotype in the Piétrain breed and 50% of Landrace individuals with the heterozygous RYR1Nn genotype The mutant RYR1n allele was found in animals of the Piétrain breed with a 100% frequency. Heterozygous genotype RYR1Nn was found in pigs of Poltava Meat (10%), Landrace (50%), Myrhorod (15%) Ukrainian Spotted Steppe (50%). Homozygous RYR1NN genotype was detected in all Large White, Vietnamese Pot-bellied, and Ukrainian Meat pigs studied, indicating the absence of stress syndrome. It has been established that pigs of the Great White, Vietnamese Pot-bellied, and Ukrainian Meat breeds are preferred for breeding for biomedical purposes. Breeding of Piétrain pigs and any combinations with this breed makes it impossible to use such animals for biomedical experimental work.