Abstract
A new effective splice variants typing based on multiplex allele-specific dual-primer RT-PCR assay was developed in a single tube for the rapid detection of the exon 7 splice variant of the BF gene. With 2 pairs of primers, one pair was used for amplifying cDNA fragments containing exon 7 of the BF gene, the other does not contain exon 7 of the BF gene. The templates were amplified in one tube and the type of splice variants was determined by the length of products to be extended and by analysis of nucleotide sequences of these BFs. Results obtained for all samples showed 100% accuracy compared to those obtained with a semi-nested PCR (snPCR) assay of 100% accuracy, but which need two round PCR assay. The dual-primer RT-PCR assay was more rapid and easy to operate than the snPCR assay. Key words: BF gene (chicken MHC class I gene), Marek’s disease (MD), splice variants, resistance, chickens.
Highlights
The Major Histocompatibility Complex (MHC) with particular traits across all jawed vertebrates came from antigens of intracellular or extracellular to the present circulating T-cells and have an integral effect on immune systems of innate and adaptive (Kelly and Trowsdale, 2019)
BL and BF genes are involved in resistance against viral, bacterial and protozoal diseases in chicken (Dawkins and Lloyd, 2019; Kaufman, 2018; Miller and Taylor, 2016; Psifidi et al, 2016)
Dual-primer reverse transcription-polymerase chain reaction (RT-PCR) detection for sensitivity showed that a 174-bp product and a 141-bp product from all dilutions but 10-6 fold dilution of the cDNA mixtures of the Xiayan chickens homozygous D12 and A5 were obtained (Figure 1)
Summary
The Major Histocompatibility Complex (MHC) with particular traits across all jawed vertebrates (two glycoproteins of primary sorts binding peptides) came from antigens of intracellular or extracellular to the present circulating T-cells and have an integral effect on immune systems of innate and adaptive (Kelly and Trowsdale, 2019). The chicken MHC on chromosome 16 have long been referred to as a gene region which makes remarkable contribution in genetic resistance to some epidemic diseases, consisting of two regions, the polymorphic MHC-Y region and the MHC-B region.
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