F.-Q. Lin

The novel HLA-A*24:02 variant, HLA-A*24:02:56, identified by sequencing in a Chinese individual

HLA‐A*24:02:56 differs from A*24:02:01:01 by one nucleotide change at position 795 from C to T.

Sequencing of the novel HLA-A*02:01:72 allele in a Chinese hematopoietic stem cell donor

HLA‐A*02:01:72 has 1 synonymous nucleotide change from HLA‐A*02:01:01:01 where 591 G is changed to A.

Identification of a novel allele, HLA-A*24:02:50, by sequence-based typing.

HLA‐A*24:02:50 has one synonymous nucleotide change from HLA‐A*24:02:01:01 at nucleotide 429 (codon 119 aspartic acid).

A novel HLA allele, HLA-B*55:50, was identified by sequence-based typing

HLA-B*55:50 has one nucleotide change from HLA-B*55:02:01 where 169 R is changed to L.

Identification of a novel HLA allele, HLA‐A*11:101, in a platelet donor from China

HLA-A*11:101 has one nucleotide change from HLA-A*11:02:01 where 120 Q is changed to R.

The novel HLA‐B allele, HLA‐B*07:110, was identified by sequencing genomic DNA

The human leukocyte antigen-B (HLA-B) gene is the most polymorphic locus of the HLA system, with more than 3000 alleles identified up to July 2013 according to the latest version of IMGT/HLA Database (Release 3.13.0) (1). Here we describe a new HLA-B allele, officially named B*07:110 found in a volunteer of platelet donor registry during routine HLA typing (2). Genomic DNA was isolated from anti-coagulated whole blood with a DNA Extraction Reagent Kit (Tiangen, Beijing, China). DNA concentration was 76 ng/μl and optical density 260/280 was 1.76. The sample was genotyped with microbeads sequence-specific oligonucleotide (SSO) assay based on Luminex platform using LABType® SSO typing kits (One Lambda Inc., Los Angeles, CA) following the manufacturer’s protocol for HLA class I (HLA-A, and -B). But the HLA-B locus hybridization probe reaction patterns did not match any known HLA-B alleles or allelic combinations, repeated typing also showed the same results, so the presence of a new HLA-B allele was suspected. To ascertain the alleles of HLA-B in the sample, DNA sequence-based typing (SBT) was then carried out. HLAB locus was amplified from exon 1 through exon 7 by SBTexcellerator® kit (Genome Diagnostics B.V., Arnhem, the Netherlands) and LongRange PCR kit (Qiagen, Hilden, Germany). Polymerase chain reaction (PCR) products were confirmed by agarose gel electrophoresis, and then treated with ExoI/shrimp alkaline phosphatase (SAP) prior to sequencing to degrade unincorporated primers and to dephosphorylize residual nucleotides. Exons 2–4 were sequenced in both directions using SBTexcellerator® sequence primer (Genome Diagnostics B.V.), Big Dye® Terminator v3.1 Reaction Kit (Applied Biosystems, Torrance, CA) and running on ABI PRISM® 3730xl Genetic Analyzer (Applied Biosystems, Foster City, CA). Sequences were analyzed with sbtengine® software. The obtained heterozygous sequence showed the presence of a novel allele, as the nucleotide (nt) sequence did not match with any known allelic combination. To separate the two alleles and to determine which allele was novel, the sample was amplified with group-specific sequencing primer (GSSP), which was indicated by the analysis software. The sequence result showed that one allele was B*44:03:01 , and the other one was a new HLA-B*07 variant. The novel allele has been officially named HLA-B*07:110 . As shown in Figure 1, B*07:110 has two nts change from B*07:02:01 at nt 226 and nt 228 where A→G (codon 76 ATA→GTG) and resulting in a coding change, 76 I is changed to V. The name B*07:110 has been officially assigned by the World Health Organization (WHO) Nomenclature Committee for Factors of the HLA system. This follows the agreed policy that, subject to the conditions stated in the most recent Nomenclature Report, names will be assigned

Detection of the new HLA-B*08:57 allele in a voluntary platelet donor

HLA-B*08:57 has 1 nt change from HLA-B*08:01:01 at nt 108 where G→C (codon 36 ATG→ATC).

Identification of a novel HLA allele, HLA-A*01:127, in a donor from China

HLA‐A*01:127 has one nucleotide change from HLA‐A*01:25 where 158 Valine is changed to Alanine.

Identification of c.238 A>G (p.Arg80Gly) of CD59 blood group gene: A NOVEL MUTATION OF CD59 BLOOD GROUP GENE

C D59 is a cell-surface glycoprotein of approximately 20 kDa. Its function is to limit the lytic activity of the terminal complement complex C5b-9 expressed on numerous cell types, including red blood cells (RBCs). In 2014, the 35th blood group system was allocated to CD59, but very little is known about the polymorphism of the CD59 blood group in different populations. The gene encoding the CD59 blood group is located on chromosome 11p13, where it spans over 40 kbp. Alternative splicing of four to seven exons leads to multiple transcript variants, eight of them having been deposited as reference sequences in the National Center for Biotechnology Information database. All the variants, however, encode the same 128–amino acid protein. To investigate the polymorphism of the CD59 blood group, a total of 1173 healthy volunteers from the Han population of Shenyang, China, were recruited, and their CD59 encoding gene was examined. Here, we report a novel missense mutation, c.238 A > G (p.Arg80Gly), in Exon 6 of the CD59 gene (NCBI Reference Sequence: NG_008057.1) detected in a healthy Chinese donor. BRIEF METHODS

Identification of the novel HLA allele, HLA-B*40:06:07, by sequence-based typing

HLA‐B*40:06:07 has one synonymous nucleotide change from HLA‐B*40:06:01:01 at nucleotide 81 (codon 3 histidine).