BACKGROUND: In Vietnam, we observed a high incidence of carbamazepine (CBZ)-induced severe cutaneous adverse drug reactions (SCARs)-Stevens-Johnson syndrome (SJS)/toxic epidermal necrolysis (TEN), and drug-induced hypersensitivity rash with eosinophilia and systemic symptoms (DRESS). In other Asian countries, HLA-B*1502 is an established risk factor for SCARs. OBJECTIVE: The aim of our study was to determine the frequency of HLA-B*1502 in SCARs patients at a large University Medical Center in Hanoi, Vietnam. METHODS: Thirty-eight cases of SCARs caused by CBZ and 25 patients with epilepsy tolerating CBZ were enrolled in a case-controlled study. Clinical manifestations and laboratory findings were recorded for each subject. Genomic DNA was isolated using the QIAamp DNA purification system. The combination of polymerase chain reaction and sequence specific oligonucleotide probes with the Luminex 100×MAP flow cytometry dual laser system was then used to quantitate fluorescently labelled oligonucleotides attached to colour-coded microbeads. RESULTS: Cases comprised 20 SJS (52.6%), 7 TEN (18.4%), 8 overlap syndrome (21.1%), and 3 DRESS patients (7.9%). A strong association between HLA B*1502 and bullous skin reactions such as SJS/TEN and overlap was confirmed with an odds ratio (OR) of 33.78 (95% confidence interval [CI], 7.55-151.03), p < 0.0001, Sensitivity 91.4%, Specificity 76.0%, positive predictive value 84.2%, and negative predictive value 86.4%. We did not, however, observe any correlation between the presence of this allele and CBZ-induced nonbullous skin reactions (DRESS) (OR, 6.33; 95% CI, 0.48-82.74; p = 0.1592). CONCLUSION: Our results indicate the presence of HLA-B*1502 in Vietnamese is a pharmacogenetic risk factor for developing CBZ-induced SJS/TEN.
Academic Medical Centers ; Alleles ; Asian Continental Ancestry Group ; Carbamazepine ; Case-Control Studies ; Cicatrix ; DNA ; Drug-Related Side Effects and Adverse Reactions ; Eosinophilia ; Epilepsy ; Exanthema ; Flow Cytometry ; HLA-B Antigens ; Humans ; Hypersensitivity ; Incidence ; Microspheres ; Odds Ratio ; Oligonucleotide Probes ; Oligonucleotides ; Pharmacogenetics ; Polymerase Chain Reaction ; Risk Factors ; Sensitivity and Specificity ; Skin ; Vietnam

BACKGROUND: Narcolepsy is a neurologic disorder characterized by excessive daytime sleepiness, symptoms of abnormal rapid eye movement (REM) sleep, and a strong association with HLA-DRB1*1501, -DQA1*0102, and -DQB1*0602. Here, we investigated the clinico-physical characteristics of Korean patients with narcolepsy, their HLA types, and the clinical utility of high-resolution PCR with sequence-specific primers (PCR-SSP) as a simple typing method for identifying DRB1*15/16, DQA1, and DQB1 alleles. METHODS: The study population consisted of 67 consecutively enrolled patients having unexplained daytime sleepiness and diagnosed narcolepsy based on clinical and neurological findings. Clinical data and the results of the multiple sleep latency test and polysomnography were reviewed, and HLA typing was performed using both high-resolution PCR-SSP and sequence-based typing (SBT). RESULTS: The 44 narcolepsy patients with cataplexy displayed significantly higher frequencies of DRB1*1501 (Pc= 0.003), DQA1*0102 (Pc=0.001), and DQB1*0602 (Pc=0.014) than the patients without cataplexy. Among patients carrying DRB1*1501-DQB1*0602 or DQA1*0102, the frequencies of a mean REM sleep latency of less than 20 min in nocturnal polysomnography and clinical findings, including sleep paralysis and hypnagogic hallucination were significantly higher. SBT and PCR-SSP showed 100% concordance for high-resolution typing of DRB1*15/16 alleles and DQA1 and DQB1 loci. CONCLUSIONS: The clinical characteristics and somnographic findings of narcolepsy patients were associated with specific HLA alleles, including DRB1*1501, DQA1*0102, and DQB1*0602. Application of high-resolution PCR-SSP, a reliable and simple method, for both allele- and locus-specific HLA typing of DRB1*15/16, DQA1, and DQB1 would be useful for characterizing clinical status among subjects with narcolepsy.
Adolescent ; Adult ; Aged ; Alleles ; Cataplexy/genetics ; *DNA Probes, HLA ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; HLA-DQ Antigens/genetics ; HLA-DRB1 Chains/genetics ; *Histocompatibility Testing ; Humans ; Male ; Middle Aged ; Narcolepsy/*diagnosis/genetics ; Phenotype ; Polymerase Chain Reaction

The aim of this study was to investigate the factors which affect HLA typing in 311 umbilical cord blood (UCB) samples. The HLA low resolution typing of UCB samples with misinterpreted HLA types from 311 UCB samples analyzed by PCR-SSO and PCR-SSP was performed. 7 samples difficult to determine their HLA genotype were sequenced directly and the reason leading to misinterpret HLA typing was analyzed. The results indicated that 99.4% of misinterpreted samples resulted from the restriction of HLA typing method itself and 0.6% of misinterpreted samples were suspected to be contaminated with maternal blood in UCB. It is concluded that HLA typing is mainly affected by the shortcomings of oligonucleotide probe design for PCR-SSO and lack of allele specific primers of PCR-SSP.
Alleles ; Base Sequence ; DNA Primers ; Fetal Blood ; immunology ; Genotype ; HLA Antigens ; genetics ; Histocompatibility Testing ; methods ; Humans ; Oligonucleotide Probes ; Polymerase Chain Reaction ; methods

OBJECTIVETo identify HLA novel allele in Chinese Han individual.

METHODSAn unknown HLA-B allele which was similar to HLA-B*5610 was detected by polymerase chain reaction-sequence specific oligonucleotide probes(PCR-SSOP), PCR-sequence specific primer(PCR-SSP) and heterozygous sequence-based typing (SBT) in a Chinese Han individual. Its anomalous patterns suggested the possible presence of new allele. The HLA-B*56 allele was amplified separately by using allele-specific primers and sequencing exons 2-4 in both directions. The differences between the novel B*56 allele and B 5610 were identified.

RESULTSThere were 4nt changes from B*5610 in exon 3, at nt379 where C>G (codon 127 CTG>GTG, 127 Leu>Val); nt412 where A>G (codon 138 AAC>GAC, 138 Asn>Asp), nt419 where T>C and nt420 where A>C (codon 140 TTA>TCC, 140 Leu>Ser). The sequence was submitted to Genbank and the accession number was EF016753.

CONCLUSIONThis allele is a novel HLA-B allele, and has been officially named HLA-B*5618 by the WHO Nomenclature Committee in September 2006.

Alleles ; China ; ethnology ; Ethnic Groups ; genetics ; Exons ; Gene Frequency ; HLA-B Antigens ; genetics ; Haplotypes ; Heterozygote ; Humans ; Male ; Oligonucleotide Probes ; genetics ; Polymerase Chain Reaction ; Sequence Analysis, DNA

OBJECTIVETo research and compare HLA-DQB1 gene frequency(GF) and polymorphism distribution between south and north population of Chinese in China.

METHODSCombining PCR-sequence specific primers(SSP) and sequence specific oligonucleotide probe(SSOP) techniques and DNA Microarray Kit for HLA-DQB1 Low Res Genotyping from Shenzhen Yi-Shengtang Biological LTD. Co. was used to type HLA-DQB1 gene polymorphisms of 700 individuals living in south China and 320 individuals in north China.

RESULTSWe inspected 10 alleles of HLA-DQB1 and got a series of comprehensive and accurate statistic data.

CONCLUSIONIt is tested that HLA-DQB1*02, 05, 0601, 0602, 0603 gene frequencies are different obviously(P<0.05) between south and north Chinese. And those data will be useful to kinds of research associated with disease relevant and anthropology research.

Alleles ; Asian Continental Ancestry Group ; genetics ; China ; ethnology ; Female ; Gene Frequency ; Genetic Variation ; HLA-DQ Antigens ; genetics ; HLA-DQ beta-Chains ; Humans ; Male ; Oligonucleotide Array Sequence Analysis ; methods ; Oligonucleotide Probes ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Sequence Analysis, DNA

OBJECTIVETo investigate, at the DNA level, the polymorphism of HLA-A, -B, -Cw genes in the Chinese of Han ethnicity in Shenyang.

METHODSHybridization with polymerase chain reaction-sequence specific oligonucleotide probes (PCR-SSOP) was used to determine HLA-A, -B and -Cw genotypes of 108 unrelated healthy individuals from a Chinese Han population. These Hans were born and living in the Shenyang area.

RESULTSThe numbers of alleles identified were 21 for HLA-A, 43 for HLA-B, and 23 for HLA-Cw. All the allele frequency distributions were consistent with the Hardy-Weinberg equilibrium.

CONCLUSIONUsing molecular method, the present authors have analyzed the characteristic of HLA I distribution in a group of indigenous Hans in Shenyang and thus have provided more accurate gene data for use in related researches.

Adolescent ; Adult ; Alleles ; China ; DNA ; analysis ; genetics ; Female ; Gene Frequency ; Genotype ; HLA-A Antigens ; genetics ; HLA-B Antigens ; genetics ; HLA-C Antigens ; genetics ; Humans ; Linkage Disequilibrium ; Male ; Middle Aged ; Oligonucleotide Probes ; Polymerase Chain Reaction ; methods ; Young Adult

Over a long period of time, studies on HLA structure and function have been the research hotspots. for it is very important to understand the essential of life science and disease mechanism. With the rapid development of molecular biology, HLA typing makes great progress. It has changed from traditional serological typing to DNA-based typing. More and more HLA genotyping methods have been developed and applied. In this essay, the author reviewed and appraised all kinds of HLA genotyping techniques and introduced two new techniques.
DNA Primers ; Genetic Techniques ; Genotype ; HLA Antigens/genetics* ; Histocompatibility Testing/methods* ; Humans ; Oligonucleotide Array Sequence Analysis ; Oligonucleotide Probes ; Polymerase Chain Reaction/methods* ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single-Stranded Conformational ; Sequence Analysis, DNA/methods*

PURPOSE: Oncogene c-erbB2 produces a transmembrane protein similar in structure to the tyrosine kinase family. Overexpression of c-erbB2 is known to lower the survival rate of breast cancer patients. c-erbB2 protein is an important antigen for tumor specific cytotoxic T lymphocytes induction that is dependent on its presentation as stably complexed with HLA-A2. In 1997, Nistico P reported low frequency of c-erbB2 proto-oncogene overexpression in HLA A2 positive breast cancer patients. And then in this study, correlation of HLA-A2 and the c-erbB2 expression was investigated in breast cancer patients. MATERIALS AND METHODS: HLA-A DNA typing by locus-specific generic PCR and by hybridization with sequence-specific oligonucleotide probes (SSOP) was performed on peripheral blood lymphocytes from 52 breast cancer patients (a PCR-SSOP typing method, involving a PCR amplification in conjunction with digoxigenin labelled sequence-specific oligonucleotide probes). To determine c-erbB2 expression, immunohistochemistry from paraffin-embedded tissues in a series of 47 patients with available tissue blocks was performed by use of rabbit anti-human c-erbB2 oncoprotein (DAKO, Glostrup, Denmark). And then we statistically analyzed the relation between the expressions of HLA-A2 and c-erbB2 in breast cancer patients. RESULTS: 29 out of 52 patients (55.8%) were HLA-A2 positive. 23.4% (11out of 47 patients) of breast cancer patients overexpressed c-erbB2. The patients with c-erbB2 overexpression showed lower estrogen receptor positivity compared to those without c-erbB2 overexpression (10.5%, vs 33.3%). HLA-A2 positive patients showed 18.5% (5/27) of overexpression and HLA-A2 negative patients showed 30.0% (6/20) of c-erbB2 overexpression (p=0.283). CONCLUSIONS: We observed no correlation between HLA-A2 and prognostic factors in breast cancer such as tumor size, axillary nodal status. However, our results showed a tendency without statistical significance between HLA-A2 and high frequency of c-erbB2 overexpression. More accumulation of patients will be needed for better conclusions.
Breast Neoplasms* ; Breast* ; Digoxigenin ; DNA Fingerprinting ; Estrogens ; HLA-A Antigens ; HLA-A2 Antigen* ; Humans ; Immunohistochemistry ; Lymphocytes ; Oligonucleotide Probes ; Oncogenes ; Polymerase Chain Reaction ; Protein-Tyrosine Kinases ; Proto-Oncogenes ; Statistics as Topic ; Survival Rate ; T-Lymphocytes, Cytotoxic