MOLECULAR GENETIC STATUS OF PIGS OF UKRAINIAN BREEDS SUITABLE FOR USE IN XENOTRANPLANTATION

Abstract

Thousands of people in Ukraine and the world need organ transplants. However, the main obstacle to the wider use of transplantation remains the shortage of donor organs and tissues. To prevent human infection with viruses during organ transplantation, pigs are carefully screened for PERV retroviruses and animals with low expression levels of PERV-A and PERV-C are selected. The article is devoted to the discussion of the results of research on the frequency of retrovirus PERV types A and C in populations of Ukrainian pig breeds. The research was carried out on samples of pigs of the following breeds: Mirgorod, Great White, Poltava meat, Ukrainian meat, Ukrainian Steppe Ryaba, Landras, Pietren, Vietnamese Vyslobryukha, Vietnamese Meishan, and wild pig. Biomaterial samples (venous blood, bristles with hair bulbs) were collected for molecular genetic analysis. Isolation of genomic DNA from the samples was carried out by the salt method and using the Chelex-100 ion exchange resin. Genotyping was performed by the method of allele-specific (PCR-SSP) multiplex polymerase chain reaction using primers complementary to regions of the PERV-C, PERV-A gene loci. A fragment of the domestic pig alpha-actin locus (α-Actin) was used as an internal PCR control. Amplification was carried out in a thermal cycler "Tertsik-2" ("DNA-technology", RF). Electrophoretic separation of amplified DNA sections in multiplex PCR format was performed in a 2% agarose gel in a tris-borate electrophoresis buffer. It was established that among the studied groups of pigs, the highest concentration of retrovirus PEVR-C was found in animals of the breeds Vietnamese Vyslobrukha (100%), Poltava meat (75%), Ukrainian steppe spotted (55%), large white (50%), landrace (50%). The largest share of PEVR-A retrovirus carrier animals is characterized by the breeds of Vietnamese Vyslobrukha (100%), Poltava meat 95%), Pietren (80%) and Ukrainian meat (73%). The largest number of individuals free from PERV-A and PERV-C retroviruses were found in experimental groups of breeds of Ukrainian steppe spotted (75 and 45%, respectively), Myrhorod (32 and 75%, respectively), landrace (75 and 50%, respectively) and wild pig. Pigs these breeds can become promising organ donors for xenotransplantation.