ObjectiveTo analyze the polymorphism of HLA-A,B,and DRB1 alleles and their haplotypes in Chinese Man population.Methods Frequencies of HLA-A,B and DRB1 alleles and haplotypes were estimated by maximum-likelihood estimation method based on the genotypes of 2183 Chinese Man bone marrow donors.ResultsA total of 18 HLA-A alleles,44 HLA-B alleles and 15 HLA-DRB1 alleles were detected in Man population,and the most frequent alleles were A*02,A*11,A*24,A*30,A*33,B*13,B*35,B*46,B*51,B*40(B60),B*40(B61),B*15(B62),DRB1*04,DRB1*07,DRB1*08,DRB1*09,DRB1*11,DRB1*12,DRB1*13,DRB1*14 and DRB1*15.A*30-B*13,A*02-DRB1*15,B*13-DRB1*07 and A*30-B*13-DRB1*07 were the most frequent haplotypes in Man population for A-B,A-DRB1,B-DRB1 and A-B-DRB1 haplotype,respectively.The number of haplotypes with frequency ≥ 0.01 for A-B,A-DRB1 and B-DRB1 haplotype was 31,24 and 27,respectively,and ≥ 0.005 for A-B-DRB1 haplotype was 32.There were 14 in A-B,3 in A-DRB1,14 in B-DRB1 and 38 in A-B-DRB1 haploypes that showed strong linkage disequilibrium with ALD≥0.40.ConclusionsThe distribution of HLA-A,B and DRB1 alleles and haplotypes in Man population was similar to that in Northern Chinese Han population.

Objective To study the molecular polymorphism and the distribution of HLA-B27 subtypes in southern Chinese Han patients with Ankylosing Spondylitis and healthy controls.Methods A total of 46 samples form southern Chinese Han patients with AS and 80 non-related blood samples from healthy peripheral blood stem cell donors with B27-positive identified by rSSO Lumminex flow array assay were subjected to sequencing analysis of exon 2 ,3 and 4 of HLA-B gene by the sequence-based typing,the purified products of sequencing reaction were electrophoresed on ABI 3730 DNA sequencer and the designation of HLA-B27 allele was accomplished using the Assign3.5 software. The ambiguities and the detected "rare" alleles were confirmed using the PCR-SSP commercial kit. Results In the 46 B27-positive patients with the diagnosis of AS,four alleles,namely B2704,B2705,B2707 and B2724 were determined. The frequencies for these four alleles were 82.98%(39/47),12.77%(6/47),2.13%(1/47) and 2.13%(1/47),respectively. In the 80 B27-positive control individuals,seven B27 related alleles were identified. The frequency for the two dominant subtype B2704 and B2705 were 57.32%(47/82) and 26.83%(22/82),respectively. Both the B2706 and B2707 were observed 5 times with a frequency of 6.10%(5/82),three alleles B2703,B2715 and B2724 were detected only once with a frequency of 1.22%(1/82).Conclusion Our study shows that HLA-B2704 and B2705 were the predominant subtypes in normal healthy controls,however,B2704 was the predominant subtype for the AS group in southern Chinese Han patients.

BACKGROUND:Many articles concerned novel al eles reported in China and outside China, but some al eles were detected lately. After that, these al eles were tested again. Because frequencies of these al eles are very low, few relevant articles are reported, so these al eles are ignored easily. OBJECTIVE:To test and identify four human leukocyte antigen rare al eles that patients and donors carries. METHODS:Genomic DNA was extracted automatical y from blood samples using quick DNA purified kit, typed by human leukocyte antigen locus commercial sequence-based typing kits and confirmed by sequence-specific primers high-resolution kits. RESULTS AND CONCLUSION:Four detected rare al eles are B*27:15, B*51:39, C*07:66 and C*08:22. The four above-mentioned human leukocyte antigen rare al eles defined by National Marrow Donor Program are not rare in China. The facts prove that C*08:22 which was defined a rare al ele by National Marrow Donor Program before is a common al ele in Chinese Han nationality.

le full matching for unrelated donor-receipt pairs. However, HLA-Cw mismatching at antigen level could no longer be ignored. The results indieated that HLA-Cw genotyping should be incorporated into future CMDP unrelated marrow donor routine HLA test.

BACKGROUND:As the sequencing technology has been widely used and high-resolution confirmation of organ transplant matching has been gradualy developed, new human leukocyte antigen (HLA) aleles are emerging. However, the gene frequency of some genes cannot be calculated accurately, and there are rare reports. These genes are often ignored, and it is easy to misjudge their genotypes only according to gene frequency. OBJECTIVE:To test and analyze a rare alele, HLA-C*08:99, from a volunteer donor of hematopoietic stem cel transplantation. METHODS: Genomic DNA was extracted automaticaly from the blood sample by using quick DNA purified kit and amplified by HLA-C locus commercial sequence-based typing kit. The purified PCR product was utilized as the DNA template in the sequencing reaction, and six direct sequencing reactions of PCR product covering exons 2, 3 and 4 in both directions were performed using commercial kit. Four direct sequencing reactions of PCR product covering exon 5 in both directions, exon 6 in forward direction and exon 7 in reverse direction were performed using in-house BigDye terminator cycle sequencing reaction kit. Sequencing reaction products purified by ethanol/sodium acetate/ ethylenediaminetetraacetic acid method were sequenced by ABI PrismTM3730 DNA Sequencer. RESULTS AND CONCLUSION:The alele assignment was analyzed with Assign-SBT 3.6+ software, and the sample HLA-C typing result was C*07:04, 08:99. Increasing the sequencing analysis at exons 5, 6 and 7 of HLA-C locus wil help to make clear the ambiguous SBT result and improve the accuracy of HLA-C typing when it is necessary, which shows important significance in clinical tissue matching. Cite this article:Wang DM, Zou HY, Nie DM, Gao SQ, Wang F.Sequencing analysis of a rare human leukocyte antigen, C*08:99, from a volunteer donor of hematopoietic stem cel transplantation. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):102-106.

AIM: To identify human leukocyte antigen (HLA)-DRB11454 allele and HLA-DRB1 exon 3 sequence information in the Chinese population, which is significant for organ transplantation, cell transplantation, and human genetics.METHODS: Polymerase chain reaction sequence-based typing (PCR-SBT) was used to identify HLA-DRB1 alleles from 58 donor-recipient individuals who would undergo haemopoietic stem cell transplantation. Medium to high resolution polymerase chain reaction-reverse sequence specific oligonucleotide probe (PCR-RSSOP) was used to identify HLA-DRB1 alleles from 1 268 healthy donors from Guangdong province. The some ambiguous results of HLA-DRB114-associated alleles were confirmed by high resolution polymerase chain reaction-sequence-specific primer typing (PCR-SSP).RESULTS: HLA-DRB11403, 1406, 1410, 1412, 1418, 1425 and 1454 alleles were detected in 1 268 healthy donors.HLA-DRB11454 was confirmed in 8 ambiguous results of HLA-DRB11401/1434/1454 alleles, and HLA-DRB11454 was one of common alleles of HLA-DRB114 allele group in Guangdong population. HLA-DRB114 exon 3 sequence information was confirmed to be polymorphic in Chinese population.CONCLUSION: HLA-DRB11454 and exon 3 of DRB1 are confirmed to be polymorphic in Chinese population, further elucidating that HLA-DRB1 axon 3 sequence information is important for Han population and some minority groups.

Objective To study the distributive characteristics of HLA-A,B,DRBI alleles and haplotypes patients with ALL in southern Chinese Han.Methods The frequencies of HLA-A,B,DRB1alleles and haplotypes were estimated by Expectation-Maximization method based on the genotypes of 572patients with ALL and 5645 unrelated health donors,and then compared by chi-square test.Results The frequencies of HLA-A33(7.15%vs 9.3%,OR=0.73,P＜0.05),B58(5.93%vs 8.75%,OR=0.64,P＜0.05),DRB1*17(5.15%vs 6.30%,OR=0.82,P＜0.05)alleles and HLA-A33-B58-DRB1*17(2.46%vs 4.14%,OB=0.35,P＜0.05)haplotype were significantly lower in ALL patient groups than that in controls.The frequencies of HLA-A3(2.1%vs 1.26%,OR=1.7,P＜0.05),B51(7.25%vs 5.78%,OR=1.3,P＜0.05)and DRB*12 (16.13%vs 12.99%,OR=1.35,P＜0.05)alleles and A2-B51-DRB1*12(1.24%vs.0.89%,OR=1.66,P＜0.05)haplotype were significantly higher in ALL patient groups than that in controls.Conclusion These results indieated that HLA-A33-B58-DRB1*17 haplotype was a associated with a diminished incidence of ALL.and HLA-A3 auele or A2-B51-DRB1*12 haplotype was weakly associated with ALL.

ObjectiveTo evaluate the effect of the timed “up and go” test (TUGT) on measuring functional mobility of stroke patients.MethodsNinety hemiparetic stroke patients participated in this study. The balance, gait speed and disability of patients were measured by Berg balance scale (BBS), maximal gait speed and functional independence measure (FIM) to find out the critical value of TUGT.ResultsA good relationship existed among TUGT and the BBS,gait speed and FIM (r=-0.926—-0.674,P<0.001).The percentage of independent walking of stroke patients whose TUGT scores <10s or>20s were 100% and 8.3%. The optimal cut off values of TUGT to predict the independent walking of patients were 15.2s, and in stroke group sensitivity and specificity of TUGT were 89.4% and 79.1%.Conclusion TUGT is a reliable instrument with adequate concurrent validity to measure the functional mobility of stroke patients.

OBJECTIVETo investigate the number and ratio of ambiguous allele combinations from human leukocyte antigen (HLA) confirmatory test by sequencing based typing for unrelated donor marrow transplantation, and to establish an efficient strategy for identifying such ambiguities.

METHODSA total of 650 donor-receipt samples were genotyped for 5 loci of the HLA gene using an Atria SBT commercial kit. Exons 2, 3 and 4 of HLA-A, -B and -C, exon 2 of HLA-DRB1 and exons 2 and 3 of HLA-DQB1 were tested by routine HLA genotyping. The ratio of usual ambiguous allele combination was calculated. The ambiguities were subjected to further confirmatory test by PCR-SSP or PCR-SBT retest at outside of the routine sequencing region.

RESULTSAmong the 650 tested samples, the ratio of ambiguity at HLA-A, B, C, DRB1 and DQB1 were 76.31% (496/650), 91.08% (592/650), 97.69% (635/650), 88.62% (576/650) and 43.38% (141/650), respectively. A total of 36 ambiguous allele combinations inside the routine sequencing region and 22 ambiguous allele combinations outside of the routine sequencing region were discovered. After removing rare alleles based on the Chinese common and well documented (CWD) Allele Table (Version 1.01), 9 ambiguous CWD allele combinations inside the routine sequencing region, including 3 located in HLA-B, HLA-C and 1 located in other three HLA loci were found. Ten ambiguous CWD allele combinations outside of the routine sequencing region, including 4 located in HLA-C, -DRB1 and 1 in HLA-A, -B respectively were determined. All samples with ambiguous CWD allele combinations could be distinguished by high-resolution PCR-SSP commercial kits or PCR SBT retest at outside of the routine sequencing region.

CONCLUSIONThe common and well documented allele combinations in sequencing-based typing at five HLA loci have been analyzed. Our strategy may provide valuable information for more efficient, low cost and accurate method for high resolution genotyping of HLA genes.

Alleles ; Genotype ; HLA Antigens ; genetics ; Humans

OBJECTIVETo investigate the genetic basis for a novel allele HLA-C*01:78.

METHODSGenomic DNA was extracted from peripheral blood using a QIAGEN quick DNA extraction kit. The regions encompassing HLA-C from exon 1 to intron 3 and intron 3 to 3'UTR were amplified and cloned using a cloning sequencing kit in order to split the two alleles apart. Selected clones were sequenced to include exons 2 to 4.

RESULTSSequencing results have indicated the HLA-C alleles of the proband to be a novel C*03:04 allele. The sequence has been submitted to GenBank (KF049216). BLAST analysis has confirmed the novel allele to have one nucleotide difference as C*01:03 at genomic nt316C>A (codon 82CGC>AGC) in exon 2, which has resulted in replacement of one amino acid (82R>S).

CONCLUSIONThe novel allele has been officially named as C*01:78 by the WHO Nomenclature Committee. The HLA allele type of the proband was therefore A*02:07, 24:02; B*40:01, 46:01; C*01:78, 03:04; DQB1*05:02, 05:02; DRB1*16:02, 16:02.

Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Exons ; Female ; HLA-C Antigens ; genetics ; Humans ; Introns ; Leukemia ; genetics ; Male ; Molecular Sequence Data ; Polymorphism, Single Nucleotide ; Sequence Analysis, DNA