DNA Sequencing as a Tissue-Typing Tool

Generation of HLA-B*1516/B*1567/B*1595 and B*1517 alleles (B15 specific group) by transpecies evolution

“…HLA-B sequences were obtained by polymerase chain reaction (PCR) using specific primers (5=BE1c, 5=BE1d, and 3=BCI3) as previously described [5,10]. PCR products were purified using EXOSAP-IT® (USB Corp., Cleveland, OH) methodology and automatically sequenced in a 3700 ABI sequencer.…”

A “silent” nucleotide substitution in exon 4 is responsible for the “alternative expression” of HLA-A*01:01:38L through aberrant splicing

“…Sequencing-based typing (SBT) of the HLA-A gene was performed as previously described [18]. Sequencing of the isolated PCR-amplified A*01 allele used a combination of sense or antisense A*01 SSP primers [19] with either sense or antisense HLA-A locus-specific primers [18].…”

“…Sequencing of the isolated PCR-amplified A*01 allele used a combination of sense or antisense A*01 SSP primers [19] with either sense or antisense HLA-A locus-specific primers [18]. DNA sequencing used Big Dye Terminator chemistry (Applied Biosystems, Foster City, CA) and sequenced fragments were separated on a 3130XL Genetic Analyzer (Applied Biosystems).…”

Polymorphism of the HLA-B*15 group of alleles is generated following 5 lineages of evolution

“…HLA-B sequences were obtained by PCR using specific primers (5= BE1c, 5= BE1d, and 3= BCI3) as previously described [31,32].…”