in the present study,9 exon-containing DNA segments of dystrophin gene with 9 sets of oligonucleotide primers by two-step multiplex polymerase chain reaction (mPCR) were amplified. Subsequently,gene analysis was perfo...in the present study,9 exon-containing DNA segments of dystrophin gene with 9 sets of oligonucleotide primers by two-step multiplex polymerase chain reaction (mPCR) were amplified. Subsequently,gene analysis was performed in 36 cases of Duchenne mascular dystroply (DMD) and 4 cases of Becker muscular dystrophy(BMD). The findings showed that 17 cases of deletion were detected by using the first 5 sets of primers with a relatively high incidence of deletion detection and 2 more cases of deletion were detected by using the remaining 4 sets of primers. The total deletion rate detected by mPCR with 9 cases of primers was 47. 5% of the patients examined,suggesting that about 79. 1% of the patients with gene deletion could be detected. Thus,as a preliminary screening, the two-step mPCR can be used in the gene diagnosis of DMD/BMD. The method is not only simple, convenient and rapid,but also free from radiosotope trouble.展开更多
In order to identify the distribution of gene types of β-thalassemia and reduce the birthrates of β-thalassemia major in Guiyang area, 1054 pregnant women and their spouses from Affiliated Hospital, Guiyang Medical ...In order to identify the distribution of gene types of β-thalassemia and reduce the birthrates of β-thalassemia major in Guiyang area, 1054 pregnant women and their spouses from Affiliated Hospital, Guiyang Medical College were screened. The positive samples were analyzed with polymerase chain reaction and reverse dot blot method (PCR-RDB). When both partners were heterozygous identified as carriers for β- thalassemia, the risk of having a fetus who was homozygous or compound heterozygous was 2.66 %; the ratio of male to female was 1/1.15. Seven types of mutation were identified. CD17 and CD41-42 were dominant among them. Among the 4 cases subject to prenatal gene diagnosis, one fetus was completely normal and 3 fetuses were diagnosed as having β-thalassemia major (1 homozygous and 2 compound heterozygous). The fetuses diagnosed as β-thalassemia major were selectively terminated within two weeks. It was concluded that the birthrate of β-thalassemia major in Guiyang area was reduced and the target of improving birth outcome and child development has been achieved.展开更多
This paper describes the application of DNA amplification in vitro with the polymerase chain reaction on the prenatal diagnosis of thalassemia syndromes and on detection of HbS and HbD Punjab genes. DNA polymerase cha...This paper describes the application of DNA amplification in vitro with the polymerase chain reaction on the prenatal diagnosis of thalassemia syndromes and on detection of HbS and HbD Punjab genes. DNA polymerase chain reaction was performed on lysed amniotic fluid cells or chorionic villus samples without prior DNA extraction and on DNA sampling from dried blood spots on filter paper blotters, α-thalassemia was prenatally diagnosed by direct analysis of amplified fetal target sequences on gel electrophoresis; and β-thalassemia was predicted by Hgi AI RFLP linkage analysis with amplified β-globin DNA. HbS and HbD Punjab genes were identified by Mst Ⅱor Eco RI mapping of the amplified β-globin DNA directly on the electrophoretic gels. The analysis of the amplified DNA does not require radioactive DNA probes and Southern hybridization. The total procedure can be completed within five hours.展开更多
Two polymorphic sites of apolipoprotein B（apoB） gene-Xbal and EcoRI were examined in a sample of 103 patients with documented coronary heart disease （CHD） and 100 healthy individuals. It was demonstrated: （1） Th...Two polymorphic sites of apolipoprotein B（apoB） gene-Xbal and EcoRI were examined in a sample of 103 patients with documented coronary heart disease （CHD） and 100 healthy individuals. It was demonstrated: （1） The frequencies of rare Xallele （presence of Xbal cutting site） and E-allele （absence of cutting site） were significantly higher in CHD patients than those in controls. It was suggested that these genetic variations were associated with CHD. （2） The patients with genotype of XXhad significantly lower展开更多
INTRODUCTIONThe point mutation rate of k-ras gene at codon 12 inpancreatic adenocarcinoma is reported to be as highas 90%,and with no mutations in normalpancreas tissues or other pancreatic disorders.Wehave detected t...INTRODUCTIONThe point mutation rate of k-ras gene at codon 12 inpancreatic adenocarcinoma is reported to be as highas 90%,and with no mutations in normalpancreas tissues or other pancreatic disorders.Wehave detected the presence of k-ras gene展开更多
Point mutation is an important molecular mechanism leading to human geneticdiseases and remains one of the focal areas of research in gene diagnosis. This notereports a simple and rapid method for the detection of kno...Point mutation is an important molecular mechanism leading to human geneticdiseases and remains one of the focal areas of research in gene diagnosis. This notereports a simple and rapid method for the detection of known mutant phenylketonuria(PKU)alleles-Multiplex Allele-specific Polymerase Chain Reaction (MASPCR) whichcan identify several point mutations at the same time.展开更多
Objective A method combining the mutagenic primers PCR and restriction enzyme digestion was designed to facilitate the detection of gene mutation in familial defective apolipoprotein B 100 R3500W. Methods A pair of p...Objective A method combining the mutagenic primers PCR and restriction enzyme digestion was designed to facilitate the detection of gene mutation in familial defective apolipoprotein B 100 R3500W. Methods A pair of primer was designed and a mismatch nucleotide was introduced in its upstream primer. A segment of target DNA including the possibly mutated nucleotide was amplified by PCR and the products were digested by restriction enzyme Nco l. To overcome the potential false negative results due to improper digestion conditions, a segment of DNA with Nco1 cut size was added as reference. Results The target sequence was successfully amplified by PCR, producing a 144 bp DNA fragment as expected. When incubated with Nco1, the enzyme could digest the DNA, producing a 114 bp segment, only if it was amplified from the mutated gene, but not from the normal allele. This difference in length of DNA could be separated by electrophoresis on a 2%agarose gel. Thus we successfully detected two carriers of heterozygous FDB R3500W in 162 hypercholesterolemic patients. Conclusions Mutagenic primers PCR can be used to detect the gene mutation of apo B 100 R3500W, two cases were detected among 162 patients with hypercholesterolemia. It suggests that this mutation is not rare in China's Mainland.展开更多
文摘in the present study,9 exon-containing DNA segments of dystrophin gene with 9 sets of oligonucleotide primers by two-step multiplex polymerase chain reaction (mPCR) were amplified. Subsequently,gene analysis was performed in 36 cases of Duchenne mascular dystroply (DMD) and 4 cases of Becker muscular dystrophy(BMD). The findings showed that 17 cases of deletion were detected by using the first 5 sets of primers with a relatively high incidence of deletion detection and 2 more cases of deletion were detected by using the remaining 4 sets of primers. The total deletion rate detected by mPCR with 9 cases of primers was 47. 5% of the patients examined,suggesting that about 79. 1% of the patients with gene deletion could be detected. Thus,as a preliminary screening, the two-step mPCR can be used in the gene diagnosis of DMD/BMD. The method is not only simple, convenient and rapid,but also free from radiosotope trouble.
文摘In order to identify the distribution of gene types of β-thalassemia and reduce the birthrates of β-thalassemia major in Guiyang area, 1054 pregnant women and their spouses from Affiliated Hospital, Guiyang Medical College were screened. The positive samples were analyzed with polymerase chain reaction and reverse dot blot method (PCR-RDB). When both partners were heterozygous identified as carriers for β- thalassemia, the risk of having a fetus who was homozygous or compound heterozygous was 2.66 %; the ratio of male to female was 1/1.15. Seven types of mutation were identified. CD17 and CD41-42 were dominant among them. Among the 4 cases subject to prenatal gene diagnosis, one fetus was completely normal and 3 fetuses were diagnosed as having β-thalassemia major (1 homozygous and 2 compound heterozygous). The fetuses diagnosed as β-thalassemia major were selectively terminated within two weeks. It was concluded that the birthrate of β-thalassemia major in Guiyang area was reduced and the target of improving birth outcome and child development has been achieved.
文摘This paper describes the application of DNA amplification in vitro with the polymerase chain reaction on the prenatal diagnosis of thalassemia syndromes and on detection of HbS and HbD Punjab genes. DNA polymerase chain reaction was performed on lysed amniotic fluid cells or chorionic villus samples without prior DNA extraction and on DNA sampling from dried blood spots on filter paper blotters, α-thalassemia was prenatally diagnosed by direct analysis of amplified fetal target sequences on gel electrophoresis; and β-thalassemia was predicted by Hgi AI RFLP linkage analysis with amplified β-globin DNA. HbS and HbD Punjab genes were identified by Mst Ⅱor Eco RI mapping of the amplified β-globin DNA directly on the electrophoretic gels. The analysis of the amplified DNA does not require radioactive DNA probes and Southern hybridization. The total procedure can be completed within five hours.
文摘Two polymorphic sites of apolipoprotein B（apoB） gene-Xbal and EcoRI were examined in a sample of 103 patients with documented coronary heart disease （CHD） and 100 healthy individuals. It was demonstrated: （1） The frequencies of rare Xallele （presence of Xbal cutting site） and E-allele （absence of cutting site） were significantly higher in CHD patients than those in controls. It was suggested that these genetic variations were associated with CHD. （2） The patients with genotype of XXhad significantly lower
基金Natural Science Foundation of Jiangsu Scientific Committee
文摘INTRODUCTIONThe point mutation rate of k-ras gene at codon 12 inpancreatic adenocarcinoma is reported to be as highas 90%,and with no mutations in normalpancreas tissues or other pancreatic disorders.Wehave detected the presence of k-ras gene
基金Project sponsored by China Medical Board (CMB)National 863 High Technology and Natural Science Foundation.
文摘Point mutation is an important molecular mechanism leading to human geneticdiseases and remains one of the focal areas of research in gene diagnosis. This notereports a simple and rapid method for the detection of known mutant phenylketonuria(PKU)alleles-Multiplex Allele-specific Polymerase Chain Reaction (MASPCR) whichcan identify several point mutations at the same time.
文摘Objective A method combining the mutagenic primers PCR and restriction enzyme digestion was designed to facilitate the detection of gene mutation in familial defective apolipoprotein B 100 R3500W. Methods A pair of primer was designed and a mismatch nucleotide was introduced in its upstream primer. A segment of target DNA including the possibly mutated nucleotide was amplified by PCR and the products were digested by restriction enzyme Nco l. To overcome the potential false negative results due to improper digestion conditions, a segment of DNA with Nco1 cut size was added as reference. Results The target sequence was successfully amplified by PCR, producing a 144 bp DNA fragment as expected. When incubated with Nco1, the enzyme could digest the DNA, producing a 114 bp segment, only if it was amplified from the mutated gene, but not from the normal allele. This difference in length of DNA could be separated by electrophoresis on a 2%agarose gel. Thus we successfully detected two carriers of heterozygous FDB R3500W in 162 hypercholesterolemic patients. Conclusions Mutagenic primers PCR can be used to detect the gene mutation of apo B 100 R3500W, two cases were detected among 162 patients with hypercholesterolemia. It suggests that this mutation is not rare in China's Mainland.
