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We report two novel HLA-C alleles, Cw*0766, and Cw*0767 identified in a Chinese voluntary stem cell donor. This sample was initially found by direct PCR-SBT with an inconclusive result. To clarify this unusual SBT result, the PCR product of HLA-C gene covering from exon 1–4 was subjected to cloning and haplotype sequencing and family study of HLA inheritance consisting of six members were confirmed. Novel Cw*0766 allele differs from the closest allele Cw*07020101 by single nucleotide change at genomic DNA nt 1651 C>A (codon 206 CTG>ATG) in exon 4, which results in an amino acid change Leu206Met. Another novel allele Cw*0767 differs from the closest allele Cw*07020103 by single nucleotide change at genomic DNA nt 930 A>T (codon 161 GAG>GTG) in exon 3, which results in an amino acid change Glu161Val. Family study demonstrated that the novel Cw*0766 allele segregating on an A*2402 B*4001 Cw*0766 DRB1*1202 haplotype was inherited from the mother of the propositus, whereas the novel Cw*0767 allele on haplotype A*1101 B*0702 Cw*0767 DRB1*0101 was inherited from the father.

The sequence of novel HLA-Cw*0124 allele differs from Cw*0103 by one nucleotide change at nt 806 from C to A, resulting in an amino acid change at codon 245 from Ala to Glu.

The full-length sequence of HLA-B*5417 differs from HLA-B*5401 only by single-nucleotide change at nt 709 where A-->C resulting in a amino acid substitution from Ile (ATC) to Val (GTC) at codon 213 in exon 4.

We report a novel human leucocyte antigen (HLA)-A allele, officially named as A*3318, which was found during high-resolution sequence-based typing in a Chinese potential hematopoietic stem cell transplantation donor. Compared with the HLA-A*330301 sequence, the A*3318 has one nucleotide substitution in exon 3: at nt 413 where A-->G resulting in an amino acid substitution from Q (CAG) to R (CGG) at codon 114.

A novel human leukocyte antigen-B (HLA-B) allele, officially named B*152703, was found during routine high-resolution sequence-based typing in a Chinese potential hematopoietic stem cell transplantation donor. Compared with the HLA-B*152701 sequence, the B*152703 has a silent substitution at position 486(G-->C) in exon 3.

HLA-DQB1*03:39 is identical to HLA-DQB1*03:03:02, except for one nucleotide change at nt 477 (C→G) in exon 3.

To establish a stable and large-scale bi-directional sequencing platform for genotyping MICA gene exons 2 to 4, and to analyze single nucleotide polymorphisms(SNP) of the region.Primers for particular alleles of MICA gene exons 2 to 5 were designed. Optimal conditions for PCR amplification and sequencing reaction were explored. A commercialized one-way sequencing kit for MICA allele was used as a parallel control. Four samples carrying a MICA *010 allele were subjected to cloning and haplotype sequencing.Results of MICA allele typing of 100 samples for a parallel control group were confirmed by the establish method. Twenty-two SNP in MICA gene exons 2 to 4 were detected in Chinese population. Two novel allelic sequences were accepted by GenBank and IMGT/HLA database and officially named as MICA*065 and MICA*066 by the WHO Nomenclature Committee. A novel SNP in MICA gene intron 3 was discovered, with allelic sequence submitted to GenBank and IMGT/HLA database.The bi-directional sequencing genotyping platform may be applied for large-scale study of MICA allelic polymorphisms, tissue typing, organ transplantation and disease research.

To identify a novel human leukocyte antigen (HLA) allele A*02:251 and analyze the sequences in Chinese population.Routine HLA-A, -B, -DRB1 high resolution genotyping for healthy Chinese donors and patients was performed with polymerase chain reaction-sequence based typing. An unknown HLA-A allele was initially detected by HLA typing in the healthy donor. Genomic DNA of the HLA-A locus in the proband was amplified, the amplified product was cloned by PMD18-T to split the two alleles, and selected clones were sequenced.The sequencing results showed that a normal A*02:06:01 and a novel A*02:251 variant allele were identified. The sequence of the novel allele has been submitted to GenBank (HM245348). Nucleotide sequence alignments with HLA-A allele from the IMGT/HLA Sequence Database showed that the novel A*02 variant allele differed from the closest allele A*02:01:01:01 by nt 383 G to C (codon 128 GAG to GAC) in exon 3, which resulted in one amino acid substitution of Glu to Asp. The HLA-A, B, C and DQB1 alleles of the healthy donor did not match with that of the patient.This novel allele is officially designated as HLA-A*02:251 by World Health Organization(WHO) Nomenclature Committee (Submission ID HWS10010755). The sequence of HLA-A locus in exon 3 is confirmed to be polymorphic in Chinese population.

A*02:01:79 is identical to A*02:01:01:01, except for one nucleotide change at nt 762 in exon 4 from C to T.

To analyze the human leukocyte antigen complex class I (-A, -B-C) and class II (-DRB1-DQB1) linked haplotypes of Guangdong Han nationality and to study the recombination events of five classical loci in the inheritance of HLA haplotypes.A total of 939 peripheral blood samples were collected from 198 families in Guangdong Han nationality who came to our center for HLA typing from 2000 August to 2009 December. HLA-(A, BDRB1) and HLA-(CDQB1) alleles were typed by low-resolution polymerase chain reaction-sequence specific oligonucleotide probes (PCR-SSO) and PCR-sequence specific primers (PCR-SSP) methods respectively. Then the recombination sites were analyzed by familial study. The samples of 52 individuals from the families with exchange recombination were analyzed by the sequence-based typing (SBT) to judge whether the recombination was interallelic or interlocus exchange.Among 543 offspring individuals of 198 families in Guangdong Han nationality, 9 individuals with HLA-A-C-B-DRB1-DQB1 linked haplotypes had a recombination rate of 1.657%. Among 9 HLA haplotypes recombined families, 3 of them were found to have a crossover between HLA-A and -Cw loci and 6 of them a crossover between HLA-B and -DRB1 loci. Four of these recombination events occurred in the most common haplotypes A*3303-Cw*0302-B*5801-DRB1*0301-DQB1*0201 of Guangdong Han nationality. Among 9 cases of recombination, 5 of them were formed by a crossover between maternal chromosomes and 4 cases a crossover between paternal chromosomes. Three individuals with an exchange between A/Cw loci were all females. Among 6 cases with an exchange between B/DRB1 loci, 5 of them were males and 1 case was female.During the inheritance, recombination of HLA linked haplotype mainly occurred between A/Cw loci and B/DRB1 loci, the recombination is related to the haplotype-specificity and sex-specificity.

HLA-Cw*0361 allele differs from Cw*030301 by one nucleotide substitution at nt 566 from T to A, resulting in an amino acid change at codon 165 from Val to Glu.

We report here the sequence of a novel human leukocyte antigen B*37 allele, B*370105, which is identical to B*370101 except for a single nucleotide substitution in exon 3 at nucleotide 558 where C>A, codon 162 GGC>GGA, no coding change.

We report here a novel human leukocyte antigen-DRB1 allele, DRB1*112802, which was identified from a Chinese individual during sequence-based typing. The new allele is identical to DRB1*112801 except for one nucleotide change at nucleotide 189 (A --> G ), codon 34 Q (CAA) --> Q (CAG), no coding change.

The genomic sequence of HLA-Cw*1222 allele is identical to Cw*120202 except for a single-nucleotide substitution at codon 87 (CAG-->CGG) resulting in an amino acid change from Gln to Arg.

HLA-Cw(*)0121 allele was identified in a Chinese patient. It is identical to Cw(*)0104 except for five nucleotides changes.

Full length sequences of this novel HLA-B*132202 allele are identical to those of HLA-B*132201 allele, except for a synonymous amino acid substitution from ACG to ACC at codon 138 in exon 3.

The novel human leukocyte antigen (HLA)-Cw*0340 allele was identified by sequence-based typing in a Chinese individual. This allele shows that the sequences of exons 2-3 of HLA-Cw*0340 are identical to those of HLA-Cw*0302, except for a nucleotide substitution that changes CTC to ATC at codon 95, resulting in an amino acid change from Leu to lle in the protein; this allele also shows that the sequences of exons 2-3 of HLA-Cw*0340 are identical to those of HLA-Cw*030401 or HLA-Cw*030403, except for a nucleotide substitution that changes TAC to TCC at codon 116, resulting in an amino acid change from Try to Ser in the protein.

We report the identification of the novel allele HLA-A*2485 that was found during routine high-resolution sequence-based typing of a Chinese bone marrow donor. The A*2485 allele has 1nt change from A*240201 at nt525 where T>A, codon 151 H>Q (CAT-->CAA).

To identify HLA novel allele in Chinese Han individuals, an unknown HLA-A allele was detected by PCR-SSP and FLOW-SSO in Chinese Han individuals. Heterozygous sequence-based typing (SBT) showed that there were 3 differences compared with database in exon 2. Its anomalous patterns suggested the possible presence of either a novel A * 30 or a novel A * 24. To separate the two alleles and to determine whether the allele is novel, the HLA-A * 30 and HLA-A * 24 alleles were amplified separately by using a commercial kit for the single allele-specific sequencing strategy, and both alleles for exons 2 - 4 were sequenced according to the manufacturer' protocol. To prepare B-lymphoblastoid cell line of the novel HLA allele by using Epstein-Barr virus-infected B-lymphoblastoid cells in the peripheral blood. The results indicated that the sequencing results showed HLA-A alleles of the sample to be HLA-A * 240201 and a new A * 30 allele. The sequences of the new A*30 were identical to those of HLA-A * 300101 except for three nucleotide changes in exon 2: at nt 121 (A--C), nt 123 (T--C) and nt 126 (A--G), resulting in an amino acid residue substitution from S (AGT) to R (CGC) at codon 17 and a synonymous substitution from G (GGA) to G (GGG) at codon 18. Immortalized B-lymphoblastoid cell line of the novel HLA-A * 3018 allele was achieved, the sequence of HLA-A * 3018 allele was submitted to GenBank and its accession number was DQ872509. In conclusion, the HLA-A * 3018 is a novel HLA-A allele and has been officially named HLA-A * 3018 by the WHO Nomenclature committee in August 2006 (HWS10004039).

The full length sequence of HLA-B*3818 differs from HLA-B*380201 at nt 660 in exon 4 (C-->A) and genomic position 2133 in intron 5 (A-->C).

The full-length sequences of exons 1-8 of this novel HLA-Cw*070205 allele are identical to those of HLA-Cw*07020101, except for one nucleotide change at nt 498 in exon 3 from C to T, which result in a synonymous amino acid substitution from ATC to ATT at codon 142 in exon 3.