1.Comparison of HLA-B27 typing methods -PCR-SSP, microlymphocytotoxicity, and flow cytometry.
Jun Soo BAE ; Young Ree KIM ; Hyun Il CHOI ; Yun Jung CHO
Korean Journal of Clinical Pathology 2000;20(2):198-203
BACKGROUND: HLA-B27 typing has long been performed by the microlymphocytotoxicity method(MCT) but the flow cytometry method(FCM) was introduced several years ago. False positive results due to the HLA-B7 cross reactive groups(CREG) were the main drawback of the serologic method. The authors performed polymerase chain reaction-sequence specific primer(PCR-SSP) test for HLA-B27 to compare the results with serologic methods. METHODS: PCR-SSP test for HLA-B27 was performed on four hundred forty one samples. Three hundred twenty eight samples were tested by MCT and one hundred thirteen samples by FCM. PCR-SSP for HLA-B27 subtyping or Amplification Refractory Mutation System-PCR(ARMS-PCR) for HLA-B typing was performed on twenty four discrepant samples. RESULTS: The concordance rate between MCT and PCR-SSP was 92.9%(305/328) and the concordance rate between FCM and PCR-SSP was 99.1%(112/113). Twenty four(5.4%) out of four hundred forty one samples showed discrepancy between serologic methods and PCR-SSP method. Fourteen out of one hundred MCT positive samples and only one out of forty FCM positive samples showed negative by PCR-SSP. Nine samples showed PCR-SSP positive and MCT negative. CONCLUSIONS: The false positive rate of MCT was quite high and there were some false positive and negative results by PCR-SSP, too. From the above findings, we suggest that FCM is the most accurate method for HLA-B27 typing in those laboratory equipped with flow cytometry.
Flow Cytometry*
;
HLA-B Antigens
;
HLA-B27 Antigen*
;
HLA-B7 Antigen
2.Sequence analysis and identification of two novel HLA alleles HLA-B*9536 and HLA-B*4612.
Yanmin HE ; Wei ZHANG ; Junjun HE ; Wei WANG ; Zhedong HAN ; Nanying CHEN ; Faming ZHU ; Lixing YAN
Chinese Journal of Medical Genetics 2009;26(6):678-680
<b>OBJECTIVEb>To identify two novel HLA alleles HLA-B*9536 and B*4612, in an individual.
<b>METHODSb>DNA was extracted from whole blood by Invitrogen DNA extraction kit. The amplification for HLA-B exons 2-4 of the proband was performed separately with allele group specific primers and the PCR products were directly sequenced for exons 2-4 in both direction.
<b>RESULTSb>There were two novel HLA-B alleles in the proband. The sequences of the two alleles have been submitted to GenBank (EU081878 and EU081879). The two alleles have been officially named as B*9536 and B*4612 by the WHO Nomenclature Committee. The sequence of exons 2-4 of HLA-B*9536 showed one nucleotide difference in exon 3 at position 544 (G to A) comparing with the closest allele B*1505, which resulted in an amino acid change from Ala to Thr at codon 158. In the HLA-B*4612 allele, there was one nucleotide change in exon 3 at position 363 (G to A), when compared to the closest allele B*4601, which lead to an amino acid change from Arg to Ser at codon 97.
<b>CONCLUSIONb>Two novel HLA-B alleles were identified in one individual and have been officially named by the WHO Nomenclature Committee.
Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; HLA-B Antigens ; genetics ; HLA-B15 Antigen ; Humans ; Male ; Molecular Sequence Data
4.HLA-B*15 gene polymorphism of registered bone marrow from donors in north Chinese Han population typed by PCR-SBT.
Jun QI ; Meng-Li LIU ; Yan ZHANG ; Sheng LIU ; Chun-Mei SHEN
Journal of Experimental Hematology 2008;16(2):429-434
To investigate the characteristics of the allele distribution of HLA-B*15 gene family in Chinese Han population and to study its influence on the selection of clinical transplantation donor, population of a 815 Han in north China from Shaanxi sub-registry of Chinese National Marrow Donor Project was randomly selected and out of them 206 HLA-B*15 positive samples according to the previous known low-resolution typing results were acquired. HLA-B*15 gene polymorphisms of above-mentioned samples and other 17 individuals were analyzed for the first time by polymerase chain reaction sequence-based typing (PCR-SBT) at high-resolution level. The structure differentiation of all HLA-B*15 alleles were analyzed by HLA three-dimensional structure modeling and software Swiss-PdbViewer. The results showed that the distribution of HLA-A, -B, -DRB1 gene of randomly selected 815 samples accorded with Hardy-Weinberg equilibrium and the gene frequency of HLA-B*15 was 0.1379. There were a total of 16 kinds of alleles of HLA-B*15 gene family to be obtained, which belonged to 7 kinds of serologic specificities. HLA-B*1501, B*1511, B*1502 and B*1518 were the major alleles with a frequency of 0.0485, 0.0215, 0.0178 and 0.0160 respectively, and the constituent ratio of their accumulated frequencies was 75.11%. The each frequency of the other 12 kinds of B*15 alleles was lower than 0.0100. Among the homozygote of 10 samples at low/medial-resolution level, there were only 4 samples to be pur sang homozygote of HLA-B*15xx, --at high-resolution level, and all the homozygote were constituted by respective dominating alleles. HLA three-dimensional structure modeling demonstrated that within the same specificity, gentle structure differentiation not only existed, such as B*1501, 1505, 1507, 1525, 1527, 1532 (each RMSD
Asian Continental Ancestry Group
;
genetics
;
China
;
ethnology
;
HLA-B Antigens
;
genetics
;
HLA-B15 Antigen
;
Humans
;
Polymerase Chain Reaction
;
methods
;
Polymorphism, Genetic
;
Tissue Donors
5.Identification of a novel HLA allele HLA-B*9526.
Yang CHEN ; Xianzhi LIU ; Xiaofeng LI ; Xu ZHANG ; Kunlian ZHANG ; Jianping LI
Chinese Journal of Medical Genetics 2009;26(6):705-708
<b>OBJECTIVEb>A novel human leukocyte antigen-B (HLA-B) allele, B*9526, was identified and analyzed by sequencing-based method in a Chinese donor.
<b>METHODSb>HLA typing was performed by PCR-sequence-specific oligonucleotide (SSO). Molecular cloning and DNA sequencing were used to identify the sequence of the potential novel allele and the difference between this new allele and other known alleles were analyzed.
<b>RESULTSb>HLA genotyping of one sample gave different results. The sequencing result showed HLA-B alleles of the proband as B*5403 and a novel allele. The nucleotide sequence of the novel allele was different from all known B alleles and harbored one nucleotide change from the closest matching allele B*1507 at nucleotide 425 (A to G) in exon 3, resulting in an amino acid change from Y (TAC) to C (TGC) at codon 142.
<b>CONCLUSIONb>A novel HLA allele was identified and officially designated as HLA-B*9526 by WHO Nomenclature Committee for Factors of the HLA System.
Alleles ; Amino Acid Sequence ; Base Sequence ; China ; Cloning, Molecular ; Genotype ; HLA-B Antigens ; genetics ; HLA-B15 Antigen ; Humans ; Male ; Molecular Sequence Data ; Sequence Analysis, DNA
6.Polymorphism of HLA-B* 40 gene family in Chinese Han population.
Zhen LI ; Shi-Zheng JIN ; Liang-Hong CHENG ; Da-Ming WANG ; Dan ZHOU ; Hong-Yan ZOU ; Guo-Guang WU
Journal of Experimental Hematology 2005;13(2):215-218
To investigate the allele distribution of HLA-B* 40 gene family in Chinese Han population and to study its influence on the selection of clinical transplantation donor, the HLA-B genetypes of 381 individuals randomly selected from Chinese National Marrow Donor Project were identified by PCR-SSO, and then all the HLA-B* 40 positive samples from the above population and the B* 40 homozygote samples received from another 1 270 registered donors were analyzed by PCR-SBT and PCR-SSP at high resolution. The results showed that the population of 381 registered donors was examined at HLA-B locus by using Hardy-Weinberg equilibrium, the gene frequency of HLA-B* 40 was 0.1692. Four different HLA-B* 40 alleles (B* 4001, B* 4002, B* 4003, B* 4006) were identified, and the serological specificity was B60 and B61 respectively. The relative frequency of each allele was 0.1192 for B* 4001, 0.0154 for B* 4002, 0.0038 for B* 4003, 0.0308 for B* 4006. The distribution of B* 40 homozygote revealed a certain regularity at high-resolution, B* 40XX (B* 4001 group), at low-resolution; B* 4001 at high resolution; B* 40XX (B* 4002 group), at low-resolution; B* 4002 or B* 4006 or heterozygote of both at high-resolution. It is concluded that in Chinese Han population, predominant allele in HLA-B* 40 gene family is B* 4001, the high-resolution typing may be recommended to use for the selection of clinical transplantation donor.
Alleles
;
Asian Continental Ancestry Group
;
genetics
;
Blood Donors
;
China
;
Gene Frequency
;
Genotype
;
HLA-B Antigens
;
genetics
;
HLA-B40 Antigen
;
Humans
;
Polymerase Chain Reaction
;
methods
;
Polymorphism, Genetic
7.Associations of HLA Alleles with Chronic Infection and Prophylaxis in Vertical Transmission of Hepatitis B Virus.
Jong Hyun KIM ; Chul Woo PYO ; Seong Suk HUR ; Yang Kyum KIM ; Dae Kyun KOH ; Jin Hee OH ; Jae Kyun HUR ; Jin Han KANG ; Soon Young PAIK ; Mun Gan RHYU ; Gum Ryong KIM ; Jee Hoon KIM ; Tai Gyu KIM
Journal of Bacteriology and Virology 2003;33(3):219-226
Perinatal transmission and infection of hepatitis B virus (HBV) in early childhood were observed in the offsprings of hepatitis B surface antigen (HBsAg)-positive mothers who had been vaccinated against HBV immediately after giving birth. This prophylaxis failure of perinatal HBV infection is likely due to the interplay of the virus and host immune response. To investigate whether the HLA polymorphism affected the outcome of the perinatal prophylaxis, HLA class I (HLA-A, B and Cw) and class II (HLA-DRB1, DQA1, DQB1 and DPB1) were typed using serology, PCR-SSOP (polymerase chain reaction-sequence specific oligonucleotide probe), and PCR-ARMS (amplification refractory modification system) methods in 22 HBeAg-positive mothers and their 10 prophylaxis-succeeded and 12 prophylaxis- failed children. The HLA types of the mothers and their children were compared with 198 HBsAg-negative healthy controls in a Korean population. HLA-B35 (relative risk=4.2, p<0.01), B51 (relative risk=3.2, p<0.02), DRB1*07 (relative risk=3.8, p<0.03), and DQA1*02 (relative risk=3.8, p<0.03) alleles were more frequent in HBeAg-positive mothers than in the controls. Also, HLA-DRB1*13 (relative risk=0.1, p<0.02) and DPB1*0401 (relative risk=0.1, p<0.02) alleles were less frequent in HBeAg-positive mothers. However, HLA alleles did not affect the outcome of the perinatal prophylaxis against HBV. These results suggest that the reported influences of some HLA alleles on the natural chronic HBV infections may not operate in the HBV infections in children received perinatal prophylaxis.
Alleles*
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Child
;
Hepatitis B Surface Antigens
;
Hepatitis B virus*
;
Hepatitis B*
;
Hepatitis*
;
HLA-B35 Antigen
;
Humans
;
Mothers
;
Parturition
8.HLA-B60 and HLA-B61 Discrimination by PCR using Sequence-specific Primers (PCR-SSP) Method.
Korean Journal of Clinical Pathology 1999;19(6):702-706
BACKGROUND: HLA-B40 is the most frequently identified HLA-B type in Koreans. Also HLA-B60 and B61 are the serologic split antigens of HLA-B40. But because of the lack of mono-specific alloantisera, cross reactivity of sera used as typing reagents, and poor antigenicity of some specific cells such as cord blood lymphocytes, discrimination between HLA-B60 and B61 has been often problematic in laboratories. In this study, authors evaluated whether the PCR-SSP method can be useful for accurate assignments of HLA-B60 and B61 or not. METHODS: Twenty-nine lymphocytes samples which were suspected as heterozygotes or homozygotes of HLA-B60 or B61 and three samples typed as HLA-B40 are selected from stored cord blood and organ transplantation donors. HLA types of these samples were defined by serologic method using a commercial typing kit. PCR that amplified exons 2 and 3 of the HLA-B gene using sequence specific primer pairs exactly matched to HLA-B60 or B61 allele making up a serological specificity was done. RESULTS: A clear discrimination between B60 and B61 was possible in all samples including 9 serologically ambiguous samples. Discrepancy between serologic typing and molecular typing was seen in three cases identified serologically as B40 positive but inable to define a split. Among three samples, two were identified as HLA-B61 and one was identified as HLA-B60. CONCLUSIONS: Molecular typing was useful in discriminating between HLA-B60 and B61. The PCR-SSP method for HLA-B60 and B61 including other cross-reactive HLA types will be helpful as a supplemental method of the serologic typing.
Alleles
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Discrimination (Psychology)*
;
Exons
;
Fetal Blood
;
Heterozygote
;
HLA-B Antigens
;
HLA-B40 Antigen
;
Homozygote
;
Humans
;
Indicators and Reagents
;
Lymphocytes
;
Molecular Typing
;
Organ Transplantation
;
Polymerase Chain Reaction*
;
Sensitivity and Specificity
;
Tissue Donors
;
Transplants
9.The associations between idiosyncratic adverse drug reactions and HLA alleles and their underlying mechanism.
Qing WANG ; Hu MEI ; Ya-Lan ZHANG ; Xian-Chao PAN ; Wen TAN ; Li CHAO
Acta Pharmaceutica Sinica 2013;48(6):799-808
With the advent of Twenty-First century, more and more genome-wide association studies (GWAS) showed that idiosyncratic adverse drug reactions (ADRs) were closely related with human leukocyte antigen (HLA) alleles, such as the associations of abacavir-HLA-B*5701, allopurinol-HLA-B*5801, and carbamazepine-HLA-B*1502, etc. To explore the mechanisms of these idiosyncratic drug reactions, hapten hypothesis, danger signal hypothesis, pharmacological interaction (P-I) concept and autoimmune mechanism are proposed. In this paper, recent GWAS studies on the HLA-mediated adverse drug reactions and underlying mechanism are reviewed in detail.
Alleles
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Allopurinol
;
adverse effects
;
Anti-HIV Agents
;
adverse effects
;
Anticonvulsants
;
adverse effects
;
Carbamazepine
;
adverse effects
;
Dideoxynucleosides
;
adverse effects
;
Drug Hypersensitivity Syndrome
;
etiology
;
immunology
;
Drug-Related Side Effects and Adverse Reactions
;
genetics
;
immunology
;
Enzyme Inhibitors
;
adverse effects
;
Genome-Wide Association Study
;
HLA Antigens
;
genetics
;
HLA-B Antigens
;
immunology
;
HLA-B15 Antigen
;
immunology
;
Humans
;
Stevens-Johnson Syndrome
;
etiology
;
immunology
10.Expression and analysis of HLA-A, B and DRB1 genes in patients with chronic myelogenous leukemia in Guangdong area.
Li WEI ; Lu-Lu XIAO ; Xiang-Yuan WU ; Qu LIN ; Ming DONG ; Jing-Yun WEN ; Xiao-Kun MA ; Fei CHONG
Journal of Experimental Hematology 2008;16(4):915-918
To study the gene polymorphism of HLA-A, B, DRB1 alleles in patients with chronic myelogenous leukemia and to explore the correlation of HLA with chronic myelogenous leukemia, the polymerase chain reaction-reverse sequence specific oligonucleotide (PCR-RSSO) was used to analyze the polymorphism of HLA-A, B, DRB1 alleles of 293 CML Patients and 406 randomized and synchronous blood donors (healthy and unrelated with patients) from Guangdong Han population. The results indicate that the gene frequency of HLA-A*24 in CML group was 15.53% lower than that of control group (22.09%, RR = 0.63, p = 0.005); the gene frequency of HLA-B*13 in CML group was 10.41% higher than that of control group (6.74%, RR = 1.68, p = 0.016). The gene frequency of HLA- DRB1*14 in CML group was 7.51% lower than that of control group (11.89%, RR = 0.58, p = 0.008). The differences were all statistically significant. It is concluded that the gene frequency of HLA-A*24, HLA- DRB1*14 in CML patients is significantly lower than normal people in Guangdong. The gene frequency of HLA-B*13 in CML patients is significantly higher than normal people in Guangdong. Further study is needed to make sure whether HLA-A*24 and HLA- DRB1*14 are protective gene markers for CML acquisition on Guangdong Chinese Han population and whether HLA-B*13 is a gene marker for CML susceptibility on this population.
Adolescent
;
Adult
;
Aged
;
Blood Donors
;
Child
;
Child, Preschool
;
China
;
Female
;
HLA-A Antigens
;
genetics
;
metabolism
;
HLA-A24 Antigen
;
HLA-B Antigens
;
genetics
;
metabolism
;
HLA-B13 Antigen
;
HLA-DR Antigens
;
genetics
;
metabolism
;
HLA-DRB1 Chains
;
Humans
;
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
;
genetics
;
immunology
;
Male
;
Middle Aged
;
Young Adult