Several colonies of pigs are used for research in medicine (NIH miniature swine, Sinclair, etc.)
because different breed of pigs may be more appropriate in different fields in research, and also it is
possible to diversify animals. Numerous inbreeding between animals, in fact, leads to a
homogeneity of genetic traits, while for special issues such as transplantation, where the
histocompatibility is critical, it is important to have the largest number of possible combinations.
The effective genotyping of the swine major histocompatibility antigens (SLA) is essential for
establishing a colony of SLA‐defined pigs (1, 2). The aim of our study is to develop and validate an
accurate and cost‐effective molecular screening for genotyping type I and type II SLA loci, having
these loci a main role in the transplantation compatibility. DNA was extracted from whole blood by
Blood purification kit using Maxwell 16 System (Promega Corp). PCR and sequencing primers
were designed in conserved regions of exon 3 of SLA1 and DQB1 genes using Primer3 software
(http:/frodo.wi.mit.edu). Amplified fragments were sequenced using BigDye Terminator
sequencing kit and 3730 DNA Analyzer (ABI). Sequences were aligned to reference alleles
available on IPDMHC Database (www.ebi.ac.uk/ipd/mhc/sla/) by Sequencer software (Gene Codes
Corp.). The protocol was developed and validated on 8 pigs, then additional 6 Swine Pathogen Free
(SPF) pigs were properly selected and genotyped to carry out inbreeding for obtaining a colony of
SLA‐homozygous pigs (genetically pure pigs). Five out of six SPF pigs resulted SLA‐identical,
showing heterozygosity for two novel alleles at SLA1 locus and homozygosity for the known
c.DQB1*0801 allele. The alleles segregation in the offspring will be analyzed to detect the genetic
phase and to attribute the nomenclature at the novel alleles. The genetically different pig resulted
homozygous for a novel allele at SLA1 gene and for the known c.DQB1*0101 allele. As the novel
SLA1 allele showed high similarity with SLA1*11 allele, the provisional name we suggest is
SLA1*11aa01. Our sequencing protocol for SLA genotyping resulted accurate, cost‐effective and
powerful, allowing to easily identify also previously undescribed alleles