Carrier detection and prenatal diagnosis of hemophilia A in a Korean population by fluorescent pcr analysis of dinucleotide repeats in intron 13 and intron 22 of the factor VIII gene / 대한산부인과학회지

OBJECTIVE: To set up the methodology for fluorescent PCR analysis of intron 13 and intron 22 microsatellite polymorphisms of the factor VIII gene, and to identify the usefulness of intron 13 and intron 22 microsatellite polymorphism for the carrier detection and prenatal diagnosis of hemophilia A in the Korean population. METHODS: Intron 13 and intron 22 microsatellite polymorphisms of the factor VIII gene were analyzed in 30 unrelated Korean mothers of patients with severe hemophilia A using fluorescent PCR. RESULTS: Analysis of intron 13 and intron 22 microsatellite polymorphisms of the factor VIII gene was feasible by the fluorescent-PCR method. The expected heterozygosity rates of intron 13 and intron 22 polymorphisms of the factor VIII gene were 67% and 34%, respectively. Combined analysis of intron 13 and intron 22 polymorphisms revealed heterozygous patterns in 16 (53%) of 30 mothers studied. Using linkage analysis with intron 13 and intron 22 polymorphisms, we have attempted three cases of carrier detection and one cases of prenatal diagnosis in two families of patients with severe hemophilia A. CONCLUSION: These results suggest that flourescent-PCR analysis of the intron 13 and intron 22 microsatellite polymorphisms within the factor VIII gene is very useful in the carrier detection and prenatal diagnosis of hemophilia A in the Korean population.

Carrier detection and prenatal diagnosis of hemophilia A in a Korean population by fluorescent pcr analysis of dinucleotide repeats in intron 13 and intron 22 of the factor VIII gene / 대한산부인과학회지

OBJECTIVE: To set up the methodology for fluorescent PCR analysis of intron 13 and intron 22 microsatellite polymorphisms of the factor VIII gene, and to identify the usefulness of intron 13 and intron 22 microsatellite polymorphism for the carrier detection and prenatal diagnosis of hemophilia A in the Korean population. METHODS: Intron 13 and intron 22 microsatellite polymorphisms of the factor VIII gene were analyzed in 30 unrelated Korean mothers of patients with severe hemophilia A using fluorescent PCR. RESULTS: Analysis of intron 13 and intron 22 microsatellite polymorphisms of the factor VIII gene was feasible by the fluorescent-PCR method. The expected heterozygosity rates of intron 13 and intron 22 polymorphisms of the factor VIII gene were 67% and 34%, respectively. Combined analysis of intron 13 and intron 22 polymorphisms revealed heterozygous patterns in 16 (53%) of 30 mothers studied. Using linkage analysis with intron 13 and intron 22 polymorphisms, we have attempted three cases of carrier detection and one cases of prenatal diagnosis in two families of patients with severe hemophilia A. CONCLUSION: These results suggest that flourescent-PCR analysis of the intron 13 and intron 22 microsatellite polymorphisms within the factor VIII gene is very useful in the carrier detection and prenatal diagnosis of hemophilia A in the Korean population.

The Relationship of P2Y1 ADP Receptor Polymorphisms and Ischemic Vascular Disease

BACKGROUND: The platelet ADP receptor P2Y1 plays a key role in platelet aggregation. METHODS: We tested eight sites of P2Y1 and studied the possible link between the presence of P2Y1 polymorphisms and the risk of is chemic vascular disease in a case-control study. The polymorphisms A1622G, C647G and C2259G were selected according to linkage disequilibrium. We evaluated 275 patients with is chemic cerebrovascular disease and 275 control subjects. We also evaluated 171 patients with acute myocardial infarction (AMI), 166 patients with unstable angina (UA), 173 patients with stable angina (SA) and 188 control subjects. RESULTS: For the cerebrovascular disease patients, A1622G AA, AG [odds ratio (OR), 1.170; 95% confidence interval (CI), 0.784 to 1.748] and GG (OR, 1.031; 95% CI, 0.554 to 1.918) did not show any difference between the case and control subjects. C647G CC, CG (OR, 0.995; 95% CI, 0.639 to 1.550) and GG (OR, 1.012; 95% CI, 0.450 to 2.277) did not show any difference between the case and control subjects. C2259G CC, CG (OR, 0.619; 95% CI, 0.354 to 1.082) and GG did not show any difference between the case and control subjects. For coronary artery disease patients, C2259G GG, CG (for AMI patients OR, 0.880, 95% CI, 0.384 to 2.016; for UA patients, OR, 0.885, 95% CI, 0.410 to 1.911; for SA patients, OR, 1.156, 95% CI, 0.534 to 2.501) and CC did not show any difference between AMI, UA and SA patients and each control subject. C647G GG, CG (for AMI patients OR, 1.351, 95% CI, 0.731 to 2.497; for UA patients OR, 1.292, 95% CI, 0.723 to 2.309; for SA patients OR, 0.977, 95% CI, 0.530 to 1.803) and CC (for AMI patients OR, 0.355, 95% CI, 0.093 to 1.358; for UA patients OR, 0.645, 95% CI, 0.205 to 2.028; for SA patients OR, 0.385, 95% CI, 0.113 to 1.311) did not show any difference between AMI, UA and SA patients and each control subject. A1622G AA, AG (for AMI patients OR, 1.416, 95% CI, 0.786 to 2.549; for UA patients OR, 1.079, 95% CI, 0.611 to 1.904; for SA patients OR, 0.958, 95% CI, 0.529 to 1.732) and GG (for AMI patients OR, 0.525, 95% CI, 0.195 to 1.411; for UA patients OR, 0.568, 95% CI, 0.231 to 1.401; for SA patients OR, 0.441, 95% CI, 0.169 to 1.154) did not show any difference between AMI, UA and, SA patients and the control subjects. CONCLUSION: The distribution of P2Y1 polymorphisms did not show any association with ischemic vascular disease.

Diagnosis of factor VIII gene inversion by PCR in Korean patients with hemophilia A and its application to carrier detection / 대한산부인과학회지

OBJECTIVE: To establish PCR (polymerase chain reaction) method for detecting factor VIII gene inversion (intron 22) causing hemophilia A, and to apply it to carrier detection of hemophilia A. DESIGN: A laboratory analysis MATERIALS AND METHODS: An inversion pattern of the factor VIII gene was analyzed in 130 unrelated Korean patients with hemophilia A and 26 female subjects using PCR. RESULTS: PCR analysis of the factor VIII gene for intron 22 inversion revealed that 91 patients (70%) were negative for the inversion, yielding 12 kb band by PQ primer. And all the other 39 (30%) patients who showed no amplification by PQ primer were positive for the inversion, yielding 11kb band by AQ primer. Among 113 patients with severe hemophilia A, 39 (35%) patients were positive for the inversion. Carrier detection for intron 22 inversion in 26 female subjects was performed, and revealed that 22 cases were carriers and 4 cases were normal female. CONCLUSION: This result suggests that PCR analysis of the inversion within the factor VIII gene is useful in the carrier detection of hemophilia A as well as in identifying hemophilia A patients with intron 22 inversion, in the Korean population.

CYP19 Gene Polymorphism in Patients with Endometriosis / 대한산부인과학회지

OBJECTIVE: To explore the association of the CYP19 gene polymorphism with the risk of endometriosis. METHODS: Two hundred seventy-nine women with surgically or histologically diagnosed endometriosis of stages I-IV (ASRM, 1997) were recruited, and two hundred eighteen patients with no evidence of endometriosis by laparoscopy or laparotomy served as control. We analysed the frequency and distribution of a TTTA repeat polymorphism and a 3 bp insertion (I)/deletion (D) polymorphism in intron 4 of the CYP19 gene. RESULTS: Six alleles of the CYP19 gene tetranucleotide repeat polymorphism were found in subjects: from 7 repeats to 13 repeats except 9 repeats. There was no statistically significant difference in the allele distribution of tetranucleotide repeat polymorphism in intron 4 of the CYP19 gene between patients with endometriosis and controls. Also there was no statistically significant difference in the 3 bp insertion (I)/deletion (D) polymorphism in intron 4 of the CYP19 gene between patients with endometriosis and controls. CONCLUSION: These results suggest that tetranucleotide repeat polymorphism and a 3 bp insertion (I)/deletion (D) polymorphism of the CYP19 gene are not associated with the risk endometriosis in the Korean population.

CYP1A1 Gene Polymorphism in Korean Women with Endometriosis / 대한산부인과학회지

OBJECTIVE: To explore the association of the CYP 1A1 gene polymorphism with the risk of endometriosis in a Korean population. DESIGN: Case-control study METHODS: Two-hundred fifty two Korean women with surgically or histologically diagnosed endometriosis of stage I-IV (ASRM, 1997) were recruited, and 203 women with no evidence of endometriosis served as controls. CYP1A1 gene MspI polymorphism was analyzed by polymerase chain reaction (PCR) and restriction fragment length polymorphism analysis. RESULTS: There was no significant difference in the genotype or allele distribution of CYP1A1 gene polymorphism between patients with endometriosis and controls. And when classified by stage, there was also no significant difference in the genotype and allele distribution of CYP1A1 gene MspI polymorphism between patients with stage I-II or stage III-IV endometriosis and controls. CONCLUSION: These results suggest that CYP1A1 gene MspI polymorphism is not associated with the risk of endometriosis in the Korean women.

Estrogen Receptor Dinucleotide Polymorphism in Patients with Endometriosis / 대한산부인과학회잡지

OBJECTIVE: To explore the association of the estrogen receptor dinucleotide repeat polymorphism with the risk of endometriosis. DESIGN: Case-control study. METHODS: One hundred fifty-one women with surgically or histologically diagnosed endometriosis of stages I-IV (ASRM, 1997) were recruited, and 137 patients with no evidence of endometriosis by laparoscopy or laparotomy served as control. Dinucleotide repeat polymorphism of the estrogen receptor gene was assessed by fluorescent PCR with gene scan analysis. Allele frequencies of dinucleotide repeat polymorphism of the estrogen receptor gene were evaluated. RESULTS: Fifteen alleles of the estrogen receptor dinucleotide repeat polymorphism were found in subjects: from 12 repeats to 27 repeats except 26 repeats. There was no statistically significant difference in the allele distribution of dinucleotide repeat polymorphism between patients with endometriosis and controls. However, patients with stage I or II endometriosis (n=51) showed a higher incidence of alleles with fewer (TA)n repeats (12-15 repeats) compared with controls (67.6% vs 52.9%, p=0.010, odds ratio=1.860). CONCLUSION: These results suggest that dinucleotide repeat polymorphism of the estrogen receptor gene is associated with the risk of minimal or mild endometriosis in the Korean population.

Molecular Genetic Diagnosis of Hemophilia A by Linkage Analysis of XbaI/intron 22 DNA Polymorphism Using PCR / 대한산부인과학회잡지

OBJECTIVE: To set up the methodology for PCR analysis of XbaI/intron 22 polymorphism of the factor VIII gene, and to identify the usefulness of XbaI/intron 22 polymorphism analysis for carrier detection and prenatal diagnosis of hemophilia A in the Korean population. DESIGN: A laboratory analysis. MATERIALS AND METHODS: A XbaI/intron 22 polymorphism of the factor VIII gene was analyzed in 56 unrelated Korean mothers of patients with severe hemophilia A, using polymerase chain reaction. RESULTS: Analysis of XbaI/intron 22 polymorphisms of the factor VIII gene were feasible by PCR method. The expected heterozygosity rates of XbaI/intron 22 polymorphism of the factor VIII gene were 44.8%. Analysis of XbaI/intron 22 polymorphism revealed heterozygous patterns in 22 (39.3%) of 56 mothers studied. Using linkage analysis with XbaI/intron 22 polymorphism, we have attempted one case of carrier detection and two cases of prenatal diagnosis in two families of patients with severe hemophilia A. CONCLUSION: These results suggest that PCR analysis of the XbaI/intron 22 polymorphism within the factor VIII gene is very useful in the carrier detection and prenatal diagnosis of hemophilia A in the Korean population.

Estrogen Receptor Gene PvuII and XbaI Polymorphism in Patients with Endometriosis / 대한산부인과학회잡지

OBJECTIVE: To explore the association of the estrogen receptor PvuII and XbaI polymorphism with endometriosis. METHODS: One hundred sixty women with surgically or histologically diagnosed endometriosis of stages I-IV, and 142 patients with no evidence of endometriosis by laparoscopy or laparotomy served as control. Frequency and distribution of PvuII and XbaI polymorphisms for estrogen receptor gene were evaluated. RESULTS: There was no statistically significant difference in the allele distribution of PvuII polymorphism between the patients and the controls (pp of 35%, pP of 51%, PP of 14% vs. 42%, 44%, 15%, p>0.1); or in the frequency of the positive PvuII allele (0.61 vs. 0.63, p>0.1). And no significant difference was also observed in the allele distribution of XbaI polymorphism between the patients and the controls (xx of 66%, xX of 29%, XX of 5% vs. 68%, 30%, 1%, p>0.1); or in the frequency of the positive XbaI allele (0.80 vs. 0.83, p>0.1). CONCLUSION: These results suggest that the PvuII or XbaI polymorphism of the estrogen receptor gene is not associated with the risk for endometriosis in the Korean population.

Fragile X Premutation in Patients with Idiopathic Premature Ovarian Failure / 대한산부인과학회잡지

OBJECTIVE: To explore the incidence of fragile X premutation in patients with idiopathic premature ovarian failure, particularly in the Korean population. DESIGN: A prospective study. MATERIALS AND METHODS: Eighty-three women affected by idiopathic premature ovarian failure were recruited for this study. Patient with known causes of premature ovarian failure were excluded: cytogenetic abnormalities, prior chemotherapy, prior bilateral oophorectomy. DNA was extracted from peripheral blood. Fragile X (FRAXA) premutation was evaluated by PCR amplification of and Southern blot analysis for FMR1 gene. RESULTS: The FRAXA premutation was detected in three (3.6%) out of 83 patients with idiopathic premature ovarian failure. CONCLUSION: This result suggests that fragile X premutation screening is indicated in patients with idiopathic premature ovarian failure, particularly in the Korean population.

The Association of GSTM1 and GSTT1 Null Polymorphisms with Endometriosis in a Korean Population / 대한산부인과학회잡지

OBJECTIVE: To explore the association of the phase II glutathione S-transferase mu (GSTM1), theta (GSTT1) gene null polymorphisms with endometriosis in a Korean population. METHODS: One hundred forty eight women with surgically diagnosed endometriosis of stage I-IV were recruited. And 130 patients with no evidence of endometriosis by laparoscopy or laparotomy served as controls. Genomic DNA was extracted from peripheral blood and analyzed for GSTM1, GSTT1 null polymorphisms. RESULTS: GSTM1 null genotype (GSTM1 0/0) was found more frequently in patients with endometriosis than in controls (61.5% in endometriosis vs. 49.2% in control, p=0.040). And the association was found only in moderate to severe endometriosis (stage III IV) (63.7% vs. 49.2%, p=0.027: odds ratio: 1.812), not in minimal or mild endometriosis (p=0.395). There was no significant difference between endometriosis patients and controls in the frequency of the GSTT1 null genotype (51.4% vs. 54.6%, p>0.1). CONCLUSION: These results suggest that GSTM1 null genotype is associated with the risk of endometriosis but GSTT1 null genotype is not in the Korean population.

N-acetyl Transferase 2 Gene Polymorphisms in Patients with Endometriosis / 대한산부인과학회잡지

OBJECTIVE: To explore the association of the N-acetyl transferase 2 (NAT 2) gene polymorphisms with endometriosis. METHODS: One hundred seventy-two women with surgically or histologically diagnosed endometriosis of stages I-IV, and 205 patients with no evidence of endometriosis by laparoscopy or laparotomy served as control. Genotype distribution of NAT 2 polymorphisms and frequencies of slow and fast acetylators in affected women and controls were evaluated. RESULTS: There was no statistically significant difference in the genotype distribution of NAT 2 polymorphisms and frequencies of slow and fast acetylators between the patients with endometriosis and the controls. CONCLUSION: These results suggest that N-acetyl transferase 2 gene polymorphisms is not associated with the risk for endometriosis in the Korean population.

Detection of Y Chromosome-specific Sequences in Patients with Turner Syndrome / 대한산부인과학회잡지

Existence of Y derived chromosome in Turner patients is significant due to the risk of gonadoblastoma development, but cytogenetic analysis may fail to detect low levels of Y chromosomal materials. Recent studies using PCR based methods showed higher sensitivity to detect Y-specific sequences, in patients who were Y chromosome-negative cytogenetically. In this study PCR was performed on 44 Turner patients with no Y chromosome by cytogenetic analysis to detect the SRY, AMELY, ZFY, and DYZ1 sequences. Of seven patients whose karyotypes were 45,X/46,X,+mar, three patients were positive for SRY, ZFY, and AMELY. DYZ1 sequences was negative in them. And any of SRY, ZFY, AMELY, and DYZ1 sequences was detected in the remaining 37 patients. This result shows that PCR analysis for Y-specific sequences in Turner patients, especially in patients who have marker chromosome is a significant effort.

The analysis of fetal gender and BclI polymorphism with fetal cells in maternal blood / 대한산부인과학회잡지

OBJECTIVE: We used nucleated erythrocytes in maternal blood for prenatal determination of the fetal gender as the preliminary experiment for the screening of fetal genetic status and the BclI DNA polymorphism in an attempt to clarify the origin of erythrocytes in maternal blood. METHODS: In seventeen pregnant women, venous blood was withdrawn and the nucleated erythrocytes were recovered by magnetic activated cell sorting (MACS) and immunostaining. After isolation of nucleated erythrocytes by micromanipulation, we performed nested PCR for amelogenin gene to identify the fetal gender and performed BclI DNA polymorphism to clarify the origin of erythrocytes. RESULTS: We could amplify the minute DNA in a single cell by primer extension preamplification and nested PCR of amelogenin gene in 94 (48.7%) cells and could identify the fetal gender by 58.8%. BclI DNA polymorphism revealed that the several cells, which did not reveal the specific band of Y chromosome in spite of the pregnancy of male fetuses, must be the cells from mother. CONCLUSION: Through this study, we could conclude that several nucleated erythrocytes in maternal blood circulation can originate from mother, therefore we must develop the new method to identify the nucleated erythrocyte of fetal origin. Considering that we must apply for the larger number of pregnant women to screen, the procedure was multi-step and complex. Therefore, we must design the new scheme to utilize the nucleated erythrocytes in maternal blood.

Analysis of Androgen Receptor Gene in Korean Patients with Androgen Insensitivity Syndrome / 대한산부인과학회잡지

Androgen insensitivity syndrome (AIS) is a X-linked disorder of sexual differentiation resulting from defective androgen receptor (AR) function. Androgens are secreted by the testes of 46,XY individuals, but there is loss of target organ response to the hormone. The abnormalities of AR are due to defects in the AR gene, and many mutations causing AIS have been reported since the cloning of AR gene. In this study, we analyzed the AR genes in twelve Korean patients with androgen insensitivity syndrome: 9 patients with complete AIS and 3 patients with partial AIS DNAs were isolated from patients with AIS, and the coding region of AR gene was amplified by a polymerase chain reaction using 7 pairs of primers (exon B-H). Sequence analysis of the AR gene was performed using direct sequencing and single strand conformational polymorphism (SSCP). The AR gene mutations were identified in 7 out of 12 patients: 6 of 9 patients with complete AIS, and one of 3 patients with partial AIS. Mutations found were as follows: the point mutation (ATT->ACT) at position 680 of exon D, point mutation (TGG->TGC) at position 751 of exon E, point mutation (CAA->TAA) at position 792 of exon F, point mutations (CGC->TGC, GTG->ATG) at position 855 and 866 of exon G, and the deletion of 13 nucleotides (CGTATCATTGCAT) at position 840 of exon G, respectively. To the best of our knowledge, the point mutations found in exon D, exon E, and exon F, and the deletion in exon G have not been observed before. SSCP revealed bands with abnormal mobility in 10 out of 12 patients tested. Mutations were found 5 out of these 10 patients. The other two patients showed no abnormal band on SSCP, but showed mutations by direct sequencing. In conclusion, we have demonstrated the AR gene mutations, including three novel mutations, in Korean patients with AIS, and these abnormalities might be related to the pathogenesis of androgen insensitivity syndrome.

Factor VIII Gene Inversions in Korean Patients with Severe Hemophilia A and its Application to Carrier Detection / 대한산부인과학회잡지

No abstract available.

Carrier Detection and Prenatal Diagnosis of Hemophilia A in a Korean Population by Analysis of Two Variable Dinucleotide Tandem Repeats within the Factor VIII Gene / 대한산부인과학회잡지

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.

Incidence of Fragile X Syndrome in Korean Patients with Mental Retardation / 대한산부인과학회잡지

Fragile X syndrome is the most common cause of inherited mental retardation. It accounts for 0.2% - 2.7% of patients with mental retardation, based upon the molecular genetic diagnosis. However, the exact prevalence of fragile X syndrome in Korean patients with mental retardation is unknown. We have performed cytogenetic and molecular analysis for fragile X syndrome in 212 Korean patients with mental retardation. Among them, six patients (2.8%) was identified as carrying fragile X syndrome by both cytogenetic and molecular analysis. The results by cytogenetic analysis was identical to those by molecular analysis. Cytogenetic analysis of 6 carriers (mothers of patients with proven fragile X syndrome) showed a fragile X chromosome in one patients (16.7%) while molecular analysis revealed premutation in all patients. PCR method using Klentaq1 Pfu polymerase showed the same results as those by PCR method using Exo(-) Pfu polymerase, but the former method is recommended because of its simplicity in technical aspect. These data suggest that the prevalence of fragile X syndrome in Korean patients with mental retardation is 2.8%, not significantly different from those in Caucasians.