6.3
#209
NINE NOVEL ALLELES IDENTIFIED BY LOW RESOLUTION MOLECULAR AND/OR SEROLOGICAL HLA TYPING, CONFIRMED BY SEQUENCE BASED TYPING.
Vaughan Carter FIBMS , Paul P.J. Dunn, PhD , Sarinder Day PhD , C. Jane Matthews, PhD and Gary Cavanagh MPhil . Newcastle upon Tyne United Kingdom, National Blood Service, Histocompatibility and Immunogenetics and Bristol United Kingdom, National Blood Service, H&I Reference Laboratory .

We report nine new HLA alleles identified using low resolution typing techniques. Molecular HLA typing techniques have led to the detection of several new HLA types that are undetectable using serology alone. Serology helps to identify expression variants such as null alleles.
Alleles were identified using a variety of low resolution HLA typing techniques: serology (polyclonal and monoclonal) and polymerase chain reaction using sequence specific primers (PCR-SSP) and sequence specific oligonucleotide probes (PCR-SSOP). All were later confirmed by PCR-sequence based typing (PCR-SBT). Eight of the nine were unique sequences while the other was a confirmatory sequence.

New alleles
AlleleSerology (polyclonal)Serology (monoclonal)PCR-SSPPCR-SSOPMutation eventd
B*0808NaB8 nullB8 nullB*08B*08D
B*0716aB7 nullB7B*07B*07S
B*1308aB13 nullB13 + Bw4 weakB*13B*13S
B*4026B21B21B*40NT4S
DRB5*0205NTNTDRB1*15, DRB5* negNT3S
A*6824A68A68A*01,*02,*33,*68No typeS
Cw*1209bNTNTCw*15Cw*04,*142S
A*02/32 hybridcA26/19A2 with A26/19A*02 with *26 and 19No typeR
Cw*07 variantcCw7Cw7Cw*07No typeS
NT=not tested; D=deletion; S=substitution; R=recombination; apreviously reported; bconfirmatory sequence; cin process of submission to EMBL-EBI database; dcompared with closest allele

These novel alleles highlight the benefits of using more than one low resolution typing technique - a single low resolution molecular method may miss a variant allele. Serology, in concert with DNA typing, has helped to explain the significance of mutations in these variants and full length gene sequencing provides a complete characterization of gene and protein.