Application of GPIIIa Gene Taq I Polymorphism to Determination of Carrier Status in Glanzmann’s Thrombasthenia Families of Chinese Origin

1993 ◽  
Vol 69 (01) ◽  
pp. 064-069 ◽  
Author(s):  
Changgeng Ruan ◽  
Jianming Gu ◽  
Xiaodong Wang ◽  
Xiaohong Chu ◽  
Junliang Pan
Keyword(s):  
Carrier Status ◽  
Glanzmann’S Thrombasthenia ◽  
Chinese Origin ◽  
Length Polymorphisms ◽  
Glanzmann's Thrombasthenia ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Asymptomatic Subjects ◽  
Autosomal Recessive Manner

SummaryGlanzmann’s thrombasthenia is a bleeding disorder caused by qualitative and/or quantitative defects of platelet membrane glycoprotein (GP) IIb/IIIa complex. The disease is inherited in an autosomal recessive manner. In this paper, cDNA probes were used to study restriction fragment length polymorphisms (RFLPs) in GPIIIa gene. A Taq I polymorphism was identified and this RFLP was composed of variant bands of 6.5 Kb/4.0 and 2.5 Kb with a frequency of 0.46/0.54 in Chinese population. The Taq I polymorphism was further localized by polymerase chain reaction (PCR) method to exon VIII of the GPIIIa gene. In two Glanzmann’s thrombasthenia families, the Taq I RFLP studied by both Southern blotting and PCR methods identified the defective GPIIIa gene inherited by patients, and determined the genotype of asymptomatic subjects. Analysis of this Taq I polymorphism by PCR method should be potentially useful in future for the carrier detection and prenatal diagnosis in Glanzmann’s thrombasthenia families.

Detection of the Glanzmann's Thrombasthenia Mutations in Arab and Iraqi-Jewish Patients by Polymerase Chain Reaction and Restriction Analysis of Blood or Urine Samples

1991 ◽  
Vol 66 (04) ◽  
pp. 500-504 ◽  
Author(s):  
H Peretz ◽  
U Seligsohn ◽  
E Zwang ◽  
B S Coller ◽  
P J Newman
Keyword(s):  
Polymerase Chain Reaction ◽  
Base Pair ◽  
Restriction Analysis ◽  
Urine Samples ◽  
Chain Reaction ◽  
Base Pairs ◽  
Glanzmann’S Thrombasthenia ◽  
Glanzmann's Thrombasthenia ◽  
Base Pair Deletion ◽  
Polymerase Chain

SummarySevere Glanzmann's thrombasthenia is relatively frequent in Iraqi-Jews and Arabs residing in Israel. We have recently described the mutations responsible for the disease in Iraqi-Jews – an 11 base pair deletion in exon 12 of the glycoprotein IIIa gene, and in Arabs – a 13 base pair deletion at the AG acceptor splice site of exon 4 on the glycoprotein IIb gene. In this communication we show that the Iraqi-Jewish mutation can be identified directly by polymerase chain reaction and gel electrophoresis. With specially designed oligonucleotide primers encompassing the mutation site, an 80 base pair segment amplified in healthy controls was clearly distinguished from the 69 base pair segment produced in patients. Patients from 11 unrelated Iraqi-Jewish families had the same mutation. The Arab mutation was identified by first amplifying a DNA segment consisting of 312 base pairs in controls and of 299 base pairs in patients, and then digestion by a restriction enzyme Stu-1, which recognizes a site that is absent in the mutant gene. In controls the 312 bp segment was digested into 235 and 77 bp fragments, while in patients there was no change in the size of the amplified 299 bp segment. The mutation was found in patients from 3 out of 5 unrelated Arab families. Both Iraqi-Jewish and Arab mutations were detectable in DNA extracted from blood and urine samples. The described simple methods of identifying the mutations should be useful for detection of the numerous potential carriers among the affected kindreds and for prenatal diagnosis using DNA extracted from chorionic villi samples.

scholarly journals PENENTUAN JENIS KELAMIN BURUNG KENARI (Serinus canaria) BERDASARKAN GEN Chromodomain Helicase DNA-Binding 1 (CHD1)

2020 ◽  
Vol 7 (1) ◽  
Author(s):  
Afif Muhammad Akhrom ◽  
Indarjulianto Soedarmanto ◽  
Yanuartono Yanuartono ◽  
Trini Susmiati ◽  
Alfarisa Nururrozi ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Sex Determination ◽  
Mature Female ◽  
Mature Male ◽  
Chain Reaction ◽  
Blood Samples ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Female Canaries

Phenotype determination of sex in young canaries is very low in accuracy. This study aimed to develop a genotypic sexing method in canaries. This study used 12 canaries consisting of 3 mature males, 3 mature females and 6 one-month-old canaries. Phenotypic sexing by cloacal observation was done on all birds, continued by genotypic sexing to identification CHD1 gene using polymerase chain reaction (PCR). The PCR used blood samples for mature canaries, and feather for mature and one-month-old canaries. The results of phenotypic observations showed that all mature male canaries had prominent and pointed cloaca forms, all mature females had flat and wide, whereas all one-month-old birds had a flat cloaca. The result of PCR showed a single band (500 bp) for mature male and double bands (500 bp and 300 bp) for mature female canaries. The PCR results of one-month-old canaries showed that there were one male and five females. Based on this study, it was concluded that genotypic sexing using the PCR method is effective in the sex determination of canaries.Keywords: canary, CHD1, genotype, PCR, sexing ABSTRAKPenentuan jenis kelamin burung kenari muda secara fenotip akurasinya sangat rendah. Penelitian ini bertujuan untuk menentukan jenis kelamin burung kenari secara genotip. Penelitian ini menggunakan 12 ekor burung kenari, terdiri dari 6 ekor dewasa (3 jantan, 3 betina) serta 6 ekor umur 1 bulan. Semua burung ditentukan jenis kelaminnya dengan mengamati kloaka dan identifikasi gen CHD1 menggunakan teknik polymerase chain reaction (PCR). Sampel DNA berasal dari darah dan bulu untuk burung dewasa serta bulu untuk burung umur 1 bulan. Pengamatan fenotip menunjukkan bahwa burung kenari dewasa jantan mempunyai bentuk kloaka menonjol dan runcing, dewasa betina berbentuk datar dan lebar, sedangkan semua burung umur 1 bulan mempunya bentuk kloaka datar. Hasil identifikasi gen CHD1 diperoleh adanya 1 pita gen sekitar 500 bp dari sampel darah dan bulu semua burung kenari dewasa jantan, dan 2 pita gen sekitar 500 bp dan 300 bp dari sampel semua burung kenari betina dewasa. Hasil PCR pada sampel burung umur 1 bulan menunjukkan bahwa 1 ekor jantan dan 5 ekor betina. Berdasarkan penelitian ini dapat disimpulkan bahwa penentuan jenis kelamin secara genotip menggunakan gen CHD1 dapat dilakukan pada burung kenari.

scholarly journals Identification of an abnormal gene for the GPIIIa subunit of the platelet fibrinogen receptor resulting in Glanzmann's thrombasthenia

Blood ◽  
1990 ◽  
Vol 75 (4) ◽  
pp. 881-888 ◽  
Author(s):  
PF Bray ◽  
MA Shuman
Keyword(s):  
Genetic Defect ◽  
Large Family ◽  
Northern Analysis ◽  
Specific Gene ◽  
Carrier Status ◽  
Glanzmann’S Thrombasthenia ◽  
Fibrinogen Receptor ◽  
Glanzmann's Thrombasthenia ◽  
Ecori Restriction ◽  
Adhesive Interactions

The platelet fibrinogen receptor, which is composed of glycoproteins IIb (GPIIb) and IIIa (GPIIIa), belongs to a large family of receptors that participate in a multitude of biologically important adhesive interactions. Platelets from most patients with the autosomal recessive bleeding disorder, Glanzmann's thrombasthenia, are deficient in GPIIb and GPIIIa. We have used cDNA probes to analyze the GPIIb and GPIIIa genes in four patients from three kindreds with Glanzmann's thrombasthenia. Southern analysis of their DNA was identical to that observed in normals when probed with a full-length GPIIb cDNA or a 3′ GPIIIa cDNA. However, in one family, a 5′ 2.0 kb GPIIIa cDNA identified abnormal DNA fragments in the father and two affected siblings' genes. A series of restriction digests resulting in small genomic fragments were probed with portions of the 5′ 2.0 kb GPIIIa cDNA and indicated that the abnormal sequences are flanked by normal fragments of the GPIIIa gene. To analyze further the genetic defect in this family, RNA was prepared from their platelets. Northern analysis revealed normal levels of GPIIb mRNA compared to control platelets. We were unable to identify GPIIIa mRNA of any size in the clinically affected family members. We also identified an EcoRI restriction fragment length polymorphism (RFLP) that permitted carrier status determination in the clinically unaffected siblings. These studies indicate that Glanzmann's thrombasthenia can be caused by heterogeneous defects in the GPIIIa gene. Furthermore, we have shown that platelets can be used to characterize normal and abnormal GPIIIa and GPIIb mRNA, and RFLPs may be used to determine the carrier status in some families with Glanzmann's thrombasthenia. The specific gene abnormality in this family appears to represent an example of an insertional mutation resulting in a human disease.

scholarly journals An exon 28 mutation resulting in alternative splicing of the glycoprotein IIb transcript and Glanzmann's thrombasthenia

Blood ◽  
1994 ◽  
Vol 83 (4) ◽  
pp. 1017-1023 ◽  
Author(s):  
S Iwamoto ◽  
E Nishiumi ◽  
E Kajii ◽  
S Ikemoto
Keyword(s):  
Family Study ◽  
Platelet Glycoprotein ◽  
Base Substitution ◽  
Mrna Levels ◽  
Glanzmann’S Thrombasthenia ◽  
Glanzmann's Thrombasthenia ◽  
Alternatively Spliced ◽  
Gene Defects ◽  
Polymerase Chain ◽  
Compound Heterozygotes

The alternatively spliced from mRNA of platelet glycoprotein IIb (GPIIb) with a deletion of exon 28 (GPIIb-28) has been isolated from the HEL cell cDNA library. The defective expression on the surface of DNA cotransfected COS-1 cells with GPIIb-28 and GPIIIa cDNAs was described in an earlier report. We studied siblings with Glanzmann's thrombasthenia who expressed only the GPIIb-28 mRNA in their platelets. Flow cytometry showed that the patients' platelets failed to bind GPIIb/IIIa complex-specific and GPIIb-specific monoclonal antibody. Western blotting showed that the patients' platelets had defective GPIIb and have trace amounts of GPIIIa. Sequence analysis was performed after polymerase chain amplification of the patients' GPIIb and GPIIIa mRNAs. The patients' GPIIb cDNA had a deletion of the exon 28 nucleotides. The polymerase chain reaction (PCR) from exon 27 to 29 showed that the GPIIb-28 mRNA was 3% +/- 1.6% of the normally, spliced form in control platelets, and 61% in the megakaryoblastic cell line UT- 7. The patients' platelets showed only the GPIIb-28. Family study and quantitative PCR studies showed that these patients were compound heterozygotes of two GPIIb gene defects. The father's allele is described in this report and involves skipping exon 28 secondary to a base substitution at codon Gln948, CAG-->TAG. The mothers allele appears to involve decreasing GPIIb mRNA levels in platelets. Our results indicate that the GPIIb-28 is not expressed on platelet membranes as a stable GPIIb/IIIa heterodimer or left as a monomer in platelets. Our studies confirm the previous data observed in COS-1 cells expressing recombinant GPIIb-28.

scholarly journals Identification of an abnormal gene for the GPIIIa subunit of the platelet fibrinogen receptor resulting in Glanzmann's thrombasthenia

Blood ◽  
1990 ◽  
Vol 75 (4) ◽  
pp. 881-888 ◽  
Author(s):  
PF Bray ◽  
MA Shuman
Keyword(s):  
Genetic Defect ◽  
Large Family ◽  
Northern Analysis ◽  
Specific Gene ◽  
Carrier Status ◽  
Glanzmann’S Thrombasthenia ◽  
Fibrinogen Receptor ◽  
Glanzmann's Thrombasthenia ◽  
Ecori Restriction ◽  
Adhesive Interactions

Abstract The platelet fibrinogen receptor, which is composed of glycoproteins IIb (GPIIb) and IIIa (GPIIIa), belongs to a large family of receptors that participate in a multitude of biologically important adhesive interactions. Platelets from most patients with the autosomal recessive bleeding disorder, Glanzmann's thrombasthenia, are deficient in GPIIb and GPIIIa. We have used cDNA probes to analyze the GPIIb and GPIIIa genes in four patients from three kindreds with Glanzmann's thrombasthenia. Southern analysis of their DNA was identical to that observed in normals when probed with a full-length GPIIb cDNA or a 3′ GPIIIa cDNA. However, in one family, a 5′ 2.0 kb GPIIIa cDNA identified abnormal DNA fragments in the father and two affected siblings' genes. A series of restriction digests resulting in small genomic fragments were probed with portions of the 5′ 2.0 kb GPIIIa cDNA and indicated that the abnormal sequences are flanked by normal fragments of the GPIIIa gene. To analyze further the genetic defect in this family, RNA was prepared from their platelets. Northern analysis revealed normal levels of GPIIb mRNA compared to control platelets. We were unable to identify GPIIIa mRNA of any size in the clinically affected family members. We also identified an EcoRI restriction fragment length polymorphism (RFLP) that permitted carrier status determination in the clinically unaffected siblings. These studies indicate that Glanzmann's thrombasthenia can be caused by heterogeneous defects in the GPIIIa gene. Furthermore, we have shown that platelets can be used to characterize normal and abnormal GPIIIa and GPIIb mRNA, and RFLPs may be used to determine the carrier status in some families with Glanzmann's thrombasthenia. The specific gene abnormality in this family appears to represent an example of an insertional mutation resulting in a human disease.

Future Directions for Pharmacogenetics

1993 ◽  
Vol 6 (1) ◽  
pp. 44-46
Author(s):  
Michael W. Jann ◽  
Y.W. Francis Lam
Keyword(s):  
Molecular Mechanisms ◽  
Regulatory Control ◽  
Ethnic Variation ◽  
Metabolizing Enzymes ◽  
Future Directions ◽  
Clinical Investigations ◽  
Length Polymorphisms ◽  
Polymerase Chain

The discipline of pharmacogenetics will continue to expand as scientific and clinical investigations increase our understanding of genetic variabilities in drug metabolism and response. These research efforts will include determination of molecular mechanisms for the different polymorphisms and evaluation of their clinical significance. The availability of molecular methodologies such as restriction fragment length polymorphisms analysis, polymerase chain reaction, and expression of cDNAs in cell cultures will further the investigative work in detection of normal and mutant alleles, identification of new substrates for different polymorphic metabolizing enzymes, and evaluation of mechanisms of individual susceptibility to biological disorders. Other areas such as the role of pharmacogenetics in drug development and regulatory control, in evaluation of potential drug-drug interactions, ethnic variation in polymorphic metabolism, and response, also need to be evaluated.

scholarly journals An exon 28 mutation resulting in alternative splicing of the glycoprotein IIb transcript and Glanzmann's thrombasthenia

Blood ◽  
1994 ◽  
Vol 83 (4) ◽  
pp. 1017-1023 ◽  
Author(s):  
S Iwamoto ◽  
E Nishiumi ◽  
E Kajii ◽  
S Ikemoto
Keyword(s):  
Family Study ◽  
Platelet Glycoprotein ◽  
Base Substitution ◽  
Mrna Levels ◽  
Glanzmann’S Thrombasthenia ◽  
Glanzmann's Thrombasthenia ◽  
Alternatively Spliced ◽  
Gene Defects ◽  
Polymerase Chain ◽  
Compound Heterozygotes

Abstract The alternatively spliced from mRNA of platelet glycoprotein IIb (GPIIb) with a deletion of exon 28 (GPIIb-28) has been isolated from the HEL cell cDNA library. The defective expression on the surface of DNA cotransfected COS-1 cells with GPIIb-28 and GPIIIa cDNAs was described in an earlier report. We studied siblings with Glanzmann's thrombasthenia who expressed only the GPIIb-28 mRNA in their platelets. Flow cytometry showed that the patients' platelets failed to bind GPIIb/IIIa complex-specific and GPIIb-specific monoclonal antibody. Western blotting showed that the patients' platelets had defective GPIIb and have trace amounts of GPIIIa. Sequence analysis was performed after polymerase chain amplification of the patients' GPIIb and GPIIIa mRNAs. The patients' GPIIb cDNA had a deletion of the exon 28 nucleotides. The polymerase chain reaction (PCR) from exon 27 to 29 showed that the GPIIb-28 mRNA was 3% +/- 1.6% of the normally, spliced form in control platelets, and 61% in the megakaryoblastic cell line UT- 7. The patients' platelets showed only the GPIIb-28. Family study and quantitative PCR studies showed that these patients were compound heterozygotes of two GPIIb gene defects. The father's allele is described in this report and involves skipping exon 28 secondary to a base substitution at codon Gln948, CAG-->TAG. The mothers allele appears to involve decreasing GPIIb mRNA levels in platelets. Our results indicate that the GPIIb-28 is not expressed on platelet membranes as a stable GPIIb/IIIa heterodimer or left as a monomer in platelets. Our studies confirm the previous data observed in COS-1 cells expressing recombinant GPIIb-28.

Determination of haemophilia A carrier status from hair samples using polymerase chain reaction technique

2008 ◽  
Vol 46 (3) ◽  
pp. 263-267 ◽  
Author(s):  
S. Hussain ◽  
A. Shamim ◽  
L. Vencer ◽  
A. I. Butt ◽  
R. Al-Harithy ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Technique ◽  
Carrier Status ◽  
Haemophilia A ◽  
Chain Reaction ◽  
Hair Samples ◽  
Polymerase Chain
Sign in / Sign up

Export Citation Format

Share Document