Molecular analysis of betaglobin gene mutations among thai. Beta thalassemia is an autosomal recessive condition characterized by decreased or absent synthesis of beta globin chains due to mutations in the beta globin gene hbb. Beta globin gene hbb sequencing can be used to identify hemoglobin variants and the most common beta thalassemia mutations, including beta plus and beta. This protein is also produced from a nearly identical gene called hba2. Molecular analysis of betaglobin gene mutations among thai beta thalassemia children. The effects of old and recent migration waves in the. The normal adult hemoglobin tetramer consists of two alpha chains and two beta chains. The alpha globin common mutation analysis test test code 11175 11174 for ny can detect the seven most common deletions and identify whether the deletions are in cis or in trans. Large deletions within the beta globin gene cluster may remove both gamma globin genes a. Kit and run on an applied biosystems abi prism 3xl genetic analyzer. This resource describes the genetics of beta globin and genetic abnormalities that give rise to blood disorders. The average percentage of total coverage for all countries beta globin gene mutation panel reaches up to approximately 77%, arguing that the herein described specific design of beta globin gene hrm analysis could be used for the detection of a significant portion of the mutations found in many european and asian areas. Investigation of mutations in the hbb gene using the 1,000 genomes. In adults, hemoglobin normally consists of four protein subunits.
In 15% of samples, additional pathogenic sequence variants were found with the majority in the other globin gene. From ncbi gene the gamma globin genes hbg1 and hbg2 are normally expressed in the fetal liver, spleen and bone marrow. Development of a highresolution melting approach for. In addition, alpha globin gene analysis detected a triple alpha globin locus which was inherited from the father. Hardison the genes encoding the different globin chains of hemoglobin are members of an ancient gene family.
Because these conditions are often complex, this test should always be interpreted in the context of protein studies, such as hemoglobin. In this study, the betaglobin gene was divided into 12 overlapping segments covering all regions of the gene from exons and introns table 2, fig. Beta globin gene sequencing is useful in the evaluation of beta globin chain variants and beta thalassemia. Firsttier testing for beta thalassemia or beta globin variant detection is thevp thalassemia and hemoglobinopathy evaluation or hbelc hemoglobin electrophoresis cascade, blood. Hemoglobin electrophoresis or high pressure liquid chromatography can reveal hemoglobin types and their amounts. Realtime multiplex analysis of four betathalassemia. Hbb protein is produced by the gene hbb which is located in the multigene locus of.
Home our laboratories specialty laboratories genetics molecular genetics tests genetic disease test menu by disorder alpha globin gene analysis text size aminus. The resulting gammadelta beta thalassemia is lethal in the homozygous state but in the heterozygote produces a transient but moderately severe microcytic anemia in. The globinclearhuman kit uses a novel, nonenzymatic technology that rapidly depletes 95% of the alpha and beta globin mrna from total rna preparations derived from whole blood. Diagnosing complex thalassemias where the beta globin gene and 1 or more of the other genes in the beta globin cluster have been deleted. Beta globin gene dosage analysis beta globin gene dosage analysis complements the sequencingbased beta globin mutation detection test in molecular diagnosis of beta thalassemia. The umass study used a similar gene editing protocol to target forms of beta thalassemia that involve splicing mutationserrors in bits of dna near the beta globin gene that change how the gene is read out to assemble beta globin protein. These sequence variants had previously not been identified given that current analysis workflows often stop as soon as a pathogenic sequence variants is found in one of the two gene clusters. No abnormal protein is present and diagnosis by electrophoresis relies on hemoglobin fraction percentage alterations ie, hb a2 or hb f elevations.
It includes information about naturallyoccurring human hemoglobin mutations and their. More than 250 mutations in the hbb gene have been found to cause beta thalassemia which is a blood disorder that reduces the production of hemoglobin and sickle cell anemia which causes distorting the red blood cell. Erlich, norman arnheim recent advances in recombinant dna technology have made possible the mo lecular analysis and prenatal diagnosis of. The betaglobin gene dosage analysis test test code 16346 16347 for ny can detect other deletions or duplications in this gene. Beta globin gene sequencing, blood asante lab test catalog. Expression of beta globin and the neighbouring globins in the. It detects almost all beta globin variants and the most common beta thalassemia mutations, although prevalence is ethnicitydependent. Hematopoeiesis originates in the embryonic yolk sac expressing the globin gene. The normal structure and regulation of human globin gene clusters bernard g. Alpha globin gene analysis dmc university laboratories. Beta globin hbb sequencing and deletionduplication.
A maternal blood sample is required for all prenatal specimens. Two gamma chains together with two alpha chains constitute fetal hemoglobin hbf which is normally replaced by adult hemoglobin hba at birth. Analysis for the hb s and hb c alleles are performed by direct sequencing of the region encompassing codon six of the hbb gene. Structural hemoglobinopathies or thalassemias insufficient or absent beta chain production.
Analysis of deletional hereditary persistence of fetal. A new standalone program called filldata provides a guided, interactive way to prepare experimental data for submission to the globin gene server. Structural defects may produce a second stable and visible hba2 fraction, whereas the unstable defects are undetectable by basic methods, and a dna analysis is required. Beta globin gene sequencing was performed on a child who was found to have hemoglobin ss disease. Beta globin gene dosage analysis test detail quest. Biochemical and molecular analysis of the betaglobin gene on. Betathalassemia mutations based on bioinformatics bioedit tools. Betaglobin complete quest diagnostics education center.
The hbbs gene is in linkage disequilibrium with five main haplotypes in the. And classified as holding bthalassemia major or bthalassemia intermedia based on their age at dianosis. These sequence variants had previously not been identified given that current analysis workflows often stop as soon as a pathogenic sequence variants is found in one of the two gene. Hbb hemoglobin subunit beta homo sapiens human gene ncbi. Molecular analysis of beta globin gene mutations among thai beta thalassemia children. How do i use the human beta globin gene primer in pcr. Pdf analysis of betaglobin gene mutations in a beta. Beta globin is a component subunit of a larger protein called hemoglobin, which is located inside red blood cells. Deltaglobin gene mutations complicate the diagnosis of. Which of the following statements best describes the effect of having the s allele, which codes for sicklecell anemia. Analysis of histone acetylation at the promoters of the globin gene clusters in uninduced k562 cells showed high levels of ach3 and ach4 at the promoters of the embryonic. The hba1 gene provides instructions for making a protein called alpha globin. The effects of these promoter substitutions on transcription were determined by transfecting the cloned mutant genes into hela cells on plasmids containing an sv40 transcription enhancer, and measuring the levels of. These two alpha globin genes are located close together in a region of chromosome 16 known as the alpha globin locus.
Analysis of beta globin gene variants in liuzhou area of guangxi. Supplies contents and method to study dna sequences. Jun 15, 2003 the globin gene cluster has long been a prototype for studying the regulation of multigenic loci 16. The molecular assessment of the entire gene fraction was performed and compared to the gene reference using dna starlasergene software. The alignment analysis pages under conserved regions in alignments now allow the user a choice between our old and new multiple alignments of the beta globin cluster. The hbb gene provides instructions for making a protein called beta globin. The beta globin gene dosage analysis test test code 16346 16347 for ny can detect other deletions or duplications in this gene. Rapid and sensitive assessment of globin chains for gene and. The tc change at position 114 of the beta globin gene results in a leucine to proline substitution leupro in the ghelix. The beta globin gene family on chromosome 11 lodish et.
Nuclear matrix association of the human globin locus. The normal structure and regulation of human globin gene clusters. Analysis of betaglobin gene mutations in a betathalassaemic patient and. A novel procedure for saturation mutagenesis of cloned dna was used to obtain more than 100 single base substitutions within the promoter of the mouse beta major globin gene. Clinical signs were compatible with severe microcytic anemia requiring bimonthly blood transfusions. Pdf in silico mutation analysis of human beta globin gene. Pcr primers designed to recognize the human beta globin gene. Betaglobin definition of betaglobin by medical dictionary. Enzymatic amplification of betaglobin genomic sequences. Beta globin gene sequencing, blood mayo clinic laboratories. Apr 19, 2004 secondary structures of singlestranded pcr products carrying both normal and mutated betaglobin gene sequences were determined using the mfold software version 3. It offers a list of experimental data and several types of software allowing comparison to find regions that are likely to play significant roles.
Hemoglobin diagnostic reference laboratory boston medical. The effects of these promoter substitutions on transcription were determined by transfecting the cloned mutant genes into hela cells on plasmids containing an sv40 transcription enhancer, and measuring the. Biochemical and molecular analysis of the betaglobin gene. This altered hemoglobin gives the blood a brown color and causes a bluish appearance of the skin, lips, and nails cyanosis. Detection of a single base pair mutation at 6th codon of.
Usually autosomal recessive, infrequently autosomal dominant. This is a secondtier evaluation of beta thalassemia minor, intermedia, and major, as well as beta globin variant identification. The beta globin gene family located on chromosome 11 is shown in the figure below and consists of five functional genes blue boxes and two pseudogenes diagonal lines. Mutations in the hbb gene are responsible for several serious hemoglobinopathies, such as sickle cell anemia and. Compared with conventional gene therapy via lentiviral vectormediated addition of antisickling. Epigenetic modifications and chromosome conformations of. Mutations at introns ivs1, ivs 2 may have a significant effect on the gene expression of the betaglobin gene warsy et al. A was under the transcriptional control of a single. Copy number variations in the form of deletions of.
Mutation analysis was performed by multiplex amplification refractory mutation system marms, direct dna sequencing of betaglobin gene. C source code is available directly from the server or by ftp from globin. Therefore, the duplicated beta globin cluster containing both. It is 147 amino acids long and has a molecular weight of 15,867 da. Beta globin medical definition merriamwebster medical. Bantu car, benin ben, senegal sen, cameroon cam and arabianindian arab. Provide groups of students 23 is best with a student version of the. Laboratory diagnosis of hemoglobinopathies and thalassemia. In silico analysis has been increasingly used to investigate the. Alphathalassemia is the most common human genetic disease worldwide.
Epigenetic analysis of the human alpha and betaglobin. Database of experimental results on gene expression. Globin gene server informs about human hemoglobin mutations. Consent forms for genetic testing may be found on the arup website at. Betathalassemia mutations based on bioinformatics bioedit. These primer sets may be used as an experimental control. Apr 26, 2017 sanger sequence analysis of the breakpoint spanning amplicon confirmed the ngs data. Were also an integral part of the center of excellence in sickle cell disease and are associated with the center.
Development of a highresolution melting approach for scanning. It is important to have simple and reliable methods to identify unknown or rare deletions. Both parents must be carriers of a thalassemia mutation for a pregnancy to be at risk. Betathalassemia prevalence, pathophysiology and inheritance. Full text molecular analysis of betaglobin gene mutations among. Enzymatic amplification of p globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia randall k. Combination of a triple alphaglobin gene with beta.
Analysis of betaglobin gene variants in liuzhou area of guangxi. Verification of software output was carried by visually looking at the chromatogram. This includes pcrbased sequencing of hba and hbb as well as deletionduplication deldup analysis by mlpa of both genes. It includes information about naturallyoccurring human hemoglobin mutations and their effects, experimental data related to the regulation of the beta like globin gene cluster, and software tools for comparing sequences with one another to discover regions that are likely to play significant roles. The effect depends on whether the allele occurs in its homozygous or heterozygous form, and the environment i. In methemoglobinemia, beta globin type, mutations in the hbb gene alter the beta globin protein and promote the heme iron to change from ferrous to ferric. The betaglobin gene is located on chromosome 11 and consists of three exons separated by two interconnected sequences of introns known as ivs. Takaras beta globin human primer set contains a set of pcr primers that recognizes the human beta globin gene at chromosome 11. Enzymatic amplification of pglobin genomic sequences and. The hemoglobin diagnostic reference laboratory at boston medical center provides clinical, genetic, and laboratory correlation and consultation, and is one of only three labs in the united states to conduct hemoglobin diagnostics testing.
This will help assess whether a fetus could be at risk for alphathalassemia major or hemoglobin h disease, if the partner is also an alphathalassemia carrier. Map of the beta globin gene libcal walter schroeder library. In addition, 195 cases suspected to have a globin gene mutation hb a 2. Ask them to find the protein sequence and highlight it on the gene strip.
Melting curve analysis was performed using the gene scanning software. This is likely due to the precision of crisprcas9 in correcting the. Population genetics and selection of hbs beta globin haplotypes background the sickle allele hbs of the beta globin gene hbb, which in the homozygous state gives rise to sickle cell disease scd, occurs commonly in populations of african ancestry as a result of its protective effect against. Molecular analysis of betaglobin gene mutations among. Population genetics and selection of hbs betaglobin. It includes information about naturallyoccurring human hemoglobin mutations and their effects, experimental data related to the regulation of the betalike globin gene cluster, and software tools for comparing sequences with one another to discover regions that are likely to play significant roles. For example, if one parent carries an alpha globin triplication 5 alpha gene copies and the other parent carries a beta zero mutation, the fetus could be at risk for beta thalassemia intermedia. Sickle cell hemoglobin is the result of a mutation at the sixth amino acid position of the beta. Eight different deletional hpfhthalassemia were observed at a frequency of 0. Two new methods were used to establish a rapid and highly sensitive prenatal diagnostic test for sickle cell anemia. Biochemical and molecular analysis of the beta globin gene on saudi.
In silico mutation analysis of human beta globin gene in sickle cell disease patients hira mubeen 1,3, rubab zahra naqvi 3, ammara masood 3, muhammad waseem shoaib 2. The first involves the primermediated enzymatic amplification of specific beta globin target sequences in genomic dna, resulting in the exponential increase 220,000 times of target dna copies. Please contact 866 gene info to speak with a genetic counselor before ordering this test. It detects large deletionsduplications involving the beta globin gene.
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