We have undertaken this study to identify the usefulness of two variable dinucleotide tandem repeats within the factor VIII gene for carrier detection and prenatal diagnosis of hemophilia A in the Korean population. We have analyzed these polymorphisms in 50 unrelated Korean mothers of patients with severe hemophilia A, using polymerase chain reaction. The expected heterozygosity rates of the intron 13 and intron 22 dinucleotide repeats were 56% and 40%, respectively. Analysis of the intron 13 and intron 22 dinucleotide repeats revealed heterozygous patterns in 29(58%) and 17(34%) of 50 mothers studied, respectively. The combined overall informativity of the intron 13 and intron 22 dinucleotide repeats was 68%. Using linkage analysis with the intron 13 dinucleotide repeats, we have attempted three cases of carrier detection and two cases of prenatal diagnosis in two families of patients with severe hemophilia A. Two pregnant women were diagnosed as carriers, and the other patients as non-carrier Prenatal diagnosis revealed an unaffected male in one fetus, and an unaffected female in another fetus. This data demonstrated that the analysis of the intron 13 and intron 22 dinucleotide repeats very useful in the carrier detection and prenatal diagnosis of hemophilia A in the Korean population.
Dinucleotide Repeats ; Factor VIII* ; Female ; Fetus ; Hemophilia A* ; Humans ; Introns ; Male ; Microsatellite Repeats ; Mothers ; Polymerase Chain Reaction ; Pregnant Women ; Prenatal Diagnosis* ; Tandem Repeat Sequences*

Tryptophan hydroxylase (TPH), the rate-limiting enzyme in serotonin biosynthesis, is primarily expressed in serotonergic neurons of the raphe nuclei. Simple tandem repeat polymorphisms, typically one to four nucleotides long, are tandemly repeated several times and often characterized by many alleles. To identify the presence of polymorphic repeats, we sequenced the 5'-upstream region of the mouse TPH gene. For the detection of any allelic variants, polymerase chain reaction, nonisotopic single-strand conformation polymophism, and DNA sequencing analyses of the tandem repeat sequences were performed using genomic DNA extracted from 60 ICR mice. Two dinucleotide repeats, 5'-(AC/TG)22-3' and 5'-(GT/CA)17-3', were identified at approximately -5.7 kb and -3.4 kb upstream from the transcriptional initiation site of the mouse TPH gene, respectively. Minor allelic variants, 5'-(AC/TG)21-3' and 5'-(GT/CA)18-3', were observed in heterozygous pairs from 3 of 60 and 1 of 60 ICR mice, respectively. The identification of these microsatellites in the mouse TPH promoter raises the possibility that identical and/or other polymorphic sequences might exist in the upstream region of the human TPH gene.
Alleles ; Animals ; Dinucleotide Repeats* ; DNA ; Humans ; Mice* ; Mice, Inbred ICR ; Microsatellite Repeats ; Nucleotides ; Polymerase Chain Reaction ; Raphe Nuclei ; Sequence Analysis, DNA ; Serotonergic Neurons ; Serotonin ; Tandem Repeat Sequences ; Tryptophan Hydroxylase* ; Tryptophan*

BACKGROUND: Variable-number tandem repeat (VNTR) typing is a promising method to discriminate the Mycobacterium tuberculosis isolates in molecular epidemiology. The purpose of this study is to determine the optimal VNTR combinations for discriminating isolated M. tuberculosis strains in Korea. METHODS: A total of 317 clinical isolates collected throughout Korea were genotyped by using the IS6110 restriction fragment length polymorphism (RFLP), and then analysed for the number of VNTR copies from 32 VNTR loci. RESULTS: The results of discriminatory power according to diverse combinations were as follows: 25 clusters in 83 strains were yielded from the internationally standardized 15 VNTR loci (Hunter-Gaston discriminatory index [HGDI], 0.9958), 25 clusters in 65 strains by using IS6110 RFLP (HGDI, 0.9977), 14 clusters in 32 strains in 12 hyper-variable VNTR loci (HGDI, 0.9995), 6 clusters in 13 strains in 32 VNTR loci (HDGI, 0.9998), and 7 clusters in 14 strains of both the 12 hyper-variable VNTR and IS6110 RFLP (HDGI, 0.9999). CONCLUSION: The combination of 12 hyper-variable VNTR typing can be an effective tool for genotyping Korean M. tuberculosis isolates where the Beijing strains are predominant.
Discrimination (Psychology)* ; Korea* ; Methods ; Minisatellite Repeats ; Molecular Epidemiology ; Mycobacterium tuberculosis* ; Mycobacterium* ; Polymorphism, Restriction Fragment Length ; Tandem Repeat Sequences ; Tuberculosis

OBJECTIVE: The aim of this study was to evaluate the association between a variable number of tandem repeats polymorphism at the dopamine D4 receptor gene (DRD4) and the performance of children with attention deficit hyperactivity disorder (ADHD) in a continuous performance test (CPT). METHODS: This study included 72 ADHD children (mean age=9.39+/-2.05 years) who were recruited from one child psychiatric clinic. The omission errors, commission errors, reaction time and reaction standardization in the CPT were computed. The number of 48-base pairs tandem repeats in the exon III of DRD4 was analyzed in a blind manner. RESULTS: The homozygosity of the 4-repeat allele at DRD4 was significantly associated with fewer commission errors (t=2.364, df=28.685, p=0.025) and standard deviation of reaction time (t=2.351, df=24.648, p=0.027) even after adjusting for age. The results of analyses of CPT measured values among three groups showed that the group with higher frequency of the 4-repeat allele showed a lower mean score of commission errors (F=4.268, df=2, p=0.018). CONCLUSION: These results suggest a protective role of 4-repeat allele of the DRD4 polymorphisms on commission errors in the CPT in children with ADHD.
Alleles ; Attention Deficit Disorder with Hyperactivity ; Child ; Dopamine ; Exons ; Humans ; Minisatellite Repeats ; Reaction Time ; Receptors, Dopamine D4 ; Tandem Repeat Sequences

OBJECTIVE: To understand the forensic practice of DNA profiling from minimal amount of DNA. METHODS: Serial dilutions of DNA were amplified with the PowerPlex 16 System Kit, then the genotyping of short tandem repeat(STR) was performed by ABI 377 DNA automated Sequencer. RESULTS: When the mount of DNA template was less than 250 pg, allelic drop-out apparently occurred at several loci. Other disturbed peaks, such as artefact bands and imbalanced heterozygote, also presented. CONCLUSION The anomalous results may result in incorrect genotyping. Careful and comprehensive considerations are needed to interpret the STR profile of minute DNA.
Alleles ; DNA/genetics* ; Forensic Medicine ; Genotype ; Humans ; Male ; Minisatellite Repeats/genetics* ; Sequence Analysis, DNA ; Tandem Repeat Sequences/genetics*

BACKGROUND: The evaluation of engraftment after BMT may be effectively accomplished by the analysis of genomic polymorphism, such as variable number of tandem repeat (VNTR). Discrimination potential (PD) and allelic profile of VNTR locus might be varied widely between races and geographic areas. Thus PCR-based VNTR loci to establish test panel useful in evaluating engraftment status of Korean patients after BMT were analyzed. MATERIAL AND METHODS: Thirty normal adults (15 males and 15 females), and each patient with acute lymphoblastic leukemia and severe aplastic anemia who had undergone allogeneic BMT were tested. Genomic DNAs extracted from peripheral blood lymphocytes or hair follicles were subjected to three PCR long tandem repeats (LTRs) and fifteen PCR short tandem repeats (STRs) loci analysis using silver-stain mode of detection. RESULTS: The PCR sensivity of VNTR system tested, and detection limit of minor component in mixing experiment, were 100 pg and 0.1%, respectively. The most informative marker was ACTBP2 with 93.2% of PD, and 98.0% of actual PD (APD). The most informative test panel was ACTBP2, D3S2386 and D1S1768 loci-combination with 99.6% of PD and 100.0% of combined APD. CONCLUSIONS: STRs, especially combination of ACTBP2, D3S2386, and D3S11768, were thought to be very useful screening markers for evaluating engraftment status in nonsibling allogeneic BMT. But most of allogeneic BMT are carried out between siblings, who have similar genetic marker each other, so further evaluation is need in sibling-BMT.
Adult ; Anemia, Aplastic ; Bone Marrow Transplantation* ; Bone Marrow* ; Continental Population Groups ; Discrimination (Psychology) ; DNA ; Genetic Markers ; Hair Follicle ; Humans ; Limit of Detection ; Lymphocytes ; Male ; Mass Screening ; Microsatellite Repeats ; Minisatellite Repeats ; Polymerase Chain Reaction ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Siblings ; Tandem Repeat Sequences

OBJECTIVETo screen and detect the female carriers from the DMD/BMD family members for prenatal or preimplantation genetic diagnosis.

METHODSFor the detection of DMD/BMD carriers from 27 family members in 4 families, PCR to five microsatellite markers(located in 5' terminus and intron 44, 45, 49, 50) and analysis of the short tandem repeat(STR) sequence polymorphism with the use of genescan were implemented.

RESULTSSix of the 17 female members were obligate DMD gene carriers according to the haplotype analysis of the results of the genescan, which conformed with the pedigree analysis. Besides, the authors detected five carriers and five normal females in these families with the use of the haplotype analysis only. The most polymorphic locus was STR 49, and the least was STR 50.

CONCLUSIONThe STR haploid linkage analysis using (CA)n repeats within the human dystrophin gene is a rapid,accurate, objective method and is well suited for routine use in clinical laboratories engaged in DMD/BMD linkage analysis for the detection of carrier.

Female ; Genetic Carrier Screening ; Humans ; Male ; Microsatellite Repeats ; Muscular Dystrophy, Duchenne ; genetics ; Polymerase Chain Reaction ; Tandem Repeat Sequences

OBJECTIVETo investigate genetic polymorphism of six short tandem repeat (STR) loci (D3S1358,D16S539, TH01,TPOX, CSF1PO,D7S820) in Zhejiang She ethnic population and Han population.

METHODSBy use of AmpFlSTR Cofiler kit, 6 STR loci in 108 She samples and 102 Han samples were amplified. The PCR products were electrophoresed by ABI Prism 377 sequencer; the data were analyzed by Genescan software.

RESULTSAll genotype frequencies of the 6 STR loci in She and Han ethnic groups met Hardy-Weinberg equilibrium. In the She population, the heterozygosities (H) in D3S1358,D16S539,TH01,TPOX, CSF1PO and D7S820 were 0.8028, 0.9148, 0.7522, 0.6728, 0.9123,0.8338, the exclusion of paternity(EP) were 0.4067, 0.6057, 0.4437, 0.3200, 0.5250, 0.5358, discrimination power (DP) were 0.6690, 0.7841, 0.6447, 0.5382, 0.7298, 0.7296.The combined DP, PE and polymorphism information content were 0.9991,0.9805,0.9988 respectively. There were significant differences at D3S1358, D16S539 and TPOX loci, compared with Hans.

CONCLUSIONShe population has its own STR allele distribution characteristic. The above data obtained from She population can be used not only in genetic researches and population investigation, but also in human identity and paternity testing.

China ; ethnology ; Ethnic Groups ; Gene Frequency ; Genetics, Population ; Heterozygote ; Humans ; Microsatellite Repeats ; genetics ; Polymorphism, Genetic ; Tandem Repeat Sequences ; genetics

OBJECTIVETo figure out the polymorphism of three Y-STR loci in isolated populations and explore the consanguinity of the populations with the use of Y-STR.

METHODSMale samples were selected from two isolated populations(80 and 60 males) in Zhejiang province and one open population (36 males), genescan was performed with males' DNA by genescan technology with ABI PRISM 377 sequencer at Y chromosome loci DYS388, DYS390 and DYS395.

RESULTSDYS388, DYS390, DYS395 allele counts in Yushan island population, Taohua island population and open population were 8, 9, 7, 5, 6, 7 and 6, 6, 5 respectively. Gene diversity was between 0.70-0.80 in the three populations. There was no difference in distribution of allele frequency and shared genotypes between the isolated populations and the open population by statistical test. Genetic distance is long between Taohua island population and open population, short between Yushan island population and open population, and moderate between Yushan island population and Taohua island population.

CONCLUSIONThe main allele is 129 at DYS388; 215 at DYS390; and 119 at DYS395. The distribution of allele frequency and gene diversity at DYS388, DYS390, DYS395 loci, and the shared genotypes between populations as well as the genetic distance are unable to explain the blood relationship between the isolated and open populations, suggesting the additional studies in large sample size will be necessary to use Y-STR for exploring the blood relationship between populations.

Alleles ; China ; Gene Frequency ; Genetics, Population ; Humans ; Male ; Microsatellite Repeats ; Polymorphism, Genetic ; Tandem Repeat Sequences ; genetics ; Y Chromosome ; genetics

BACKGROUND: The CBF/NF-Y enhancer region of ABO gene reported to contain 43bp minisatellite tandem repeats has been rarely reported. We describe here the relationship between minisatellite tandem repeats and ABO alleles in samples from Korean population with common ABO blood group and rare ABO subgroup. METHODS: Sixty one cases of ABO subgroup (14 A2, 12 A2B, 1 Aweak, 7 AweakB, 11 B3, 5 A1B3, 1 A1Bweak, 2 Bweak, and 8 cis-AB) and 41 cases of common ABO blood group (13 A, 6 AB, 11 B, and 11 O) were obtained from healthy donors at the Gwangju-Chonnam Red Cross Blood Center between Sep 2004 and Aug 2005. Red cells were phenotyped by standard serologic tests and genotyped by direct DNA sequencing exon 6 and 7 of the ABO gene. The minisatellite repeats were analyzed by PCR method. RESULTS: The ABO*A101 and *A102 had only one repeat, *B101, *O01 and *O02 had 4 repeats in common ABO blood group, while the *A102, *cis-AB01, and *Aw10 had only one minisatellite repeat and *A201, *A204, *B101, *Bw03, *B306, *O01, and *O02 alleles had 4 repeats and unexpectedly 3 A2 cases with *A102 had 4 repeats in the rare ABO subgroup. CONCLUSION: The minisatellite repeats found in Koreans correlate well with ABO alleles in sample common ABO phenotype, but do not completely correlate with those of ABO subgroup. We revealed here a pattern of the minisatellite repeats in various ABO subgroup in Korea.
Alleles ; Blood Donors ; Exons ; Humans ; Korea ; Minisatellite Repeats ; Phenotype ; Polymerase Chain Reaction ; Red Cross ; Sequence Analysis, DNA ; Serologic Tests ; Tandem Repeat Sequences ; Tissue Donors