Caracterização molecular e laboratorial da talassemia beta e da interação hemoglobina s/talassemia beta
Autor(a) principal: | |
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Data de Publicação: | 2010 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFRN |
Texto Completo: | https://repositorio.ufrn.br/jspui/handle/123456789/13454 |
Resumo: | Beta thalassemia arises as a consequence of the reduction (β+, β++, βsilent) or absence (β0) of beta globin chain synthesis and results from a number of mechanisms that lead to genetic defects. The inheritance of beta thalassemia is characterized by the existence of heterozygous individuals, compound heterozygotes, homozygotes and those with coinheritance of beta thalassemia allele and other thalassemias and/or hemoglobin variants. The aim of this study was to perform molecular and laboratory characterization of beta thalassemia in heterozygous and homozygous individuals and in those with coinheritance of S beta thalassemia. A total of 48 individuals were included (35 heterozygotes, 4 homozygotes and 9 S beta thalessemia carriers) referred to the Integrated Laboratory of Clinical Analyses of the Federal University of Rio Grande do Norte (UFRN) and the Hematology Ambulatory Facility of the Dalton Barbosa Cunha Hemocenter (Hemonorte Natal, Brazil). Peripheral blood samples form each patient underwent the following laboratory examinations: erythrogram, hemoglobin electrophoresis at alkaline pH, measurements of Hb A2, Fetal Hb and serum ferritin. DNA was extracted using the illustra blood genomicPrep Mini Spin Kit and molecular characterization was performed by the PCR/RFLP technique, which involves digestion with specific restriction enzymes for IVS-1 nt 1 (G®A), IVS-1 nt 6 (T®C) and codon 39 (CAG®TAG) mutations. Of the 35 heterozygotes, 37.1% showed IVS-1 nt 6 mutation, 42.9% IVS-1 nt 1 and 20% were carriers of other mutations not identified by the technique used. The four homozygous patients presented with the IVS-1 nt 6 mutation, while 66.7% of the individuals with S beta thalassemia had the IVS-1 nt 1 mutation. Codon 39 was not detected in any of the patients investigated. Of the thallasemic alleles found, 40.4% were IVS- 1 nt 1, 40.4% IVS-1 nt 6 and 19.2% were not identified. Laboratory data showed that the heterozygotes exhibited microcytosis and hypochromia, evidenced by MCV ranging from 57 to 75fL and MCH from 15.9 to 23.6 pg. Hemoglobin A2 varied between 3.7 and 7.2%. The homogygotes also showed reduced MCV and MCH and elevated HbA2.. Comparison of laboratory data between heterozygous individuals with IVS-1 nt 1 and IVS-1 nt 6 mutations showed that heterozygotes for the IVS1-1 mutation had significantly lower mean MCV and MCH (p = 0.023 and 0.007, respectively) and significantly higher hemoglobin A2 (p < 0.001) when compared to heterozygotes for the IVS-1 nt 6 mutation. PCR/RFLP was useful in identifying the presence or absence of IVS-1 nt 6, IVS-1 nt 1 and codon 39 mutations in most of the patients investigated here. This is the first study conducted in the state of Rio Grande do Norte, Brazil aimed at identifying beta thalassemia mutations and represents an important contribution to the knowledge regarding the molecular profile of beta thalassemia in our country |
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Silveira, Zama Messala Luna dahttp://lattes.cnpq.br/9321913536357216http://lattes.cnpq.br/9869053526045053Schimieguel, Dulce Martahttp://lattes.cnpq.br/6361482547166270Araújo, Aderson da Silvahttp://lattes.cnpq.br/9615780257704881Medeiros, Tereza Maria Dantas de2014-12-17T14:16:25Z2010-11-302014-12-17T14:16:25Z2010-02-25SILVEIRA, Zama Messala Luna da. Caracterização molecular e laboratorial da talassemia beta e da interação hemoglobina s/talassemia beta. 2010. 82 f. Dissertação (Mestrado em Bioanálises e Medicamentos) - Universidade Federal do Rio Grande do Norte, Natal, 2010.https://repositorio.ufrn.br/jspui/handle/123456789/13454Beta thalassemia arises as a consequence of the reduction (β+, β++, βsilent) or absence (β0) of beta globin chain synthesis and results from a number of mechanisms that lead to genetic defects. The inheritance of beta thalassemia is characterized by the existence of heterozygous individuals, compound heterozygotes, homozygotes and those with coinheritance of beta thalassemia allele and other thalassemias and/or hemoglobin variants. The aim of this study was to perform molecular and laboratory characterization of beta thalassemia in heterozygous and homozygous individuals and in those with coinheritance of S beta thalassemia. A total of 48 individuals were included (35 heterozygotes, 4 homozygotes and 9 S beta thalessemia carriers) referred to the Integrated Laboratory of Clinical Analyses of the Federal University of Rio Grande do Norte (UFRN) and the Hematology Ambulatory Facility of the Dalton Barbosa Cunha Hemocenter (Hemonorte Natal, Brazil). Peripheral blood samples form each patient underwent the following laboratory examinations: erythrogram, hemoglobin electrophoresis at alkaline pH, measurements of Hb A2, Fetal Hb and serum ferritin. DNA was extracted using the illustra blood genomicPrep Mini Spin Kit and molecular characterization was performed by the PCR/RFLP technique, which involves digestion with specific restriction enzymes for IVS-1 nt 1 (G®A), IVS-1 nt 6 (T®C) and codon 39 (CAG®TAG) mutations. Of the 35 heterozygotes, 37.1% showed IVS-1 nt 6 mutation, 42.9% IVS-1 nt 1 and 20% were carriers of other mutations not identified by the technique used. The four homozygous patients presented with the IVS-1 nt 6 mutation, while 66.7% of the individuals with S beta thalassemia had the IVS-1 nt 1 mutation. Codon 39 was not detected in any of the patients investigated. Of the thallasemic alleles found, 40.4% were IVS- 1 nt 1, 40.4% IVS-1 nt 6 and 19.2% were not identified. Laboratory data showed that the heterozygotes exhibited microcytosis and hypochromia, evidenced by MCV ranging from 57 to 75fL and MCH from 15.9 to 23.6 pg. Hemoglobin A2 varied between 3.7 and 7.2%. The homogygotes also showed reduced MCV and MCH and elevated HbA2.. Comparison of laboratory data between heterozygous individuals with IVS-1 nt 1 and IVS-1 nt 6 mutations showed that heterozygotes for the IVS1-1 mutation had significantly lower mean MCV and MCH (p = 0.023 and 0.007, respectively) and significantly higher hemoglobin A2 (p < 0.001) when compared to heterozygotes for the IVS-1 nt 6 mutation. PCR/RFLP was useful in identifying the presence or absence of IVS-1 nt 6, IVS-1 nt 1 and codon 39 mutations in most of the patients investigated here. This is the first study conducted in the state of Rio Grande do Norte, Brazil aimed at identifying beta thalassemia mutations and represents an important contribution to the knowledge regarding the molecular profile of beta thalassemia in our countryA talassemia beta ocorre devido à diminuição (β+, β++, βsilent) ou ausência (β0) de síntese de cadeias beta de globina e é decorrente de vários mecanismos que provocam o defeito genético. A herança da talassemia beta é caracterizada pela existência de indivíduos heterozigotos, heterozigoto compostos, homozigotos e portadores de interação entre o alelo talassêmico beta e outras talassemias e/ou variantes de hemoglobina. O objetivo principal deste estudo foi realizar a caracterização molecular e laboratorial em indivíduos heterozigotos e homozigotos da talassemia beta e em portadores da interação hemoglobina S/talassemia beta. Foram incluídos 48 indivíduos (35 heterozigotos, 4 homozigotos e 9 com interação HbS/talassemia beta) atendidos no Laboratório Integrado de Análises Clínicas (UFRN) e no Ambulatório de Hematologia do Hemocentro Dalton Barbosa Cunha (Hemonorte Natal/RN). As amostras de sangue periférico de cada paciente foram submetidas aos seguintes exames laboratoriais: eritrograma, eletroforese de hemoglobina em pH alcalino, dosagem das hemoglobinas A2 e Fetal, e ferritina sérica. O DNA foi extraído utilizando-se o kit blood genomicPrep Mini Spin e a caracterização molecular foi realizada através da técnica PCR/RFLP mediante digestão com enzimas de restrição específicas para as mutações IVS-1 nt 1 (G®A), IVS-1 nt 6 (T®C) e nonsense códon 39 (CAG®TAG). Dentre os 35 heterozigotos, 37,1% apresentaram a mutação IVS-1 nt 6, 42,9% a IVS-1 nt 1 e 20% eram portadores de outras mutações não identificadas com a técnica utilizada. Os quatro pacientes homozigotos apresentaram a mutação IVS-1 nt 6, enquanto 66,7% dos indivíduos com interação HbS/talassemia beta tinham a mutação IVS-1 nt 1. A códon 39 não foi detectada em nenhum dos pacientes investigados. Dentre os alelos talassêmicos encontrados, 40,4% eram IVS-1 nt 1, 40,4% eram IVS-1 nt 6 e 19,2% não foram identificados. Em relação aos dados laboratoriais, os heterozigotos apresentaram microcitose e hipocromia evidenciada pelo VCM variando de 57 a 75fL, e o HCM, entre 15,9 a 23,6 pg. A hemoglobina A2 variou de 3,7 a 7,2%. Os homozigotos também apresentaram VCM e HCM reduzidos e HbA2 elevada. Ao comparar os dados laboratoriais entre os indivíduos heterozigotos para as mutações IVS-1 nt 1 e IVS-1 nt 6 observou-se que os heterozigotos da mutação IVS1-1 apresentaram valores médios de VCM e HCM significativamente menores (p = 0,023 e 0,007, respectivamente) e hemoglobina A2 significativamente mais elevados (p < 0,001) quando comparados aos heterozigotos da mutação IVS-1 nt 6. A técnica de PCR/RFLP se mostrou útil para a identificação da presença ou ausência das mutações IVS-1 nt 6, IVS-1 nt 1 e β0 códon 39 na maioria dos pacientes investigados na pesquisa. O presente estudo é o primeiro trabalho realizado no estado do Rio Grande do Norte para identificação das mutações da talassemia beta, e constitui importante contribuição para o conhecimento do perfil molecular da talassemia beta em nosso paísCoordenação de Aperfeiçoamento de Pessoal de Nível Superiorapplication/pdfporUniversidade Federal do Rio Grande do NortePrograma de Pós-Graduação em Ciências FarmacêuticasUFRNBRBioanálises e MedicamentosTalassemia betaHemoglobinopatiasInteração hemoglobina S/talassemia betaMutaçõesPCR/RFLPβ-thalassemiaHemoglobinopathiesS-βthalassemiaMutationsPCR/RFLPCNPQ::CIENCIAS DA SAUDE::FARMACIACaracterização molecular e laboratorial da talassemia beta e da interação hemoglobina s/talassemia betainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFRNinstname:Universidade Federal do Rio Grande do Norte (UFRN)instacron:UFRNORIGINALCaracterizacaoMolecularLaboratorial_Silveira_2010.pdfapplication/pdf2983727https://repositorio.ufrn.br/bitstream/123456789/13454/1/CaracterizacaoMolecularLaboratorial_Silveira_2010.pdf7afd93b3940a02c3ef820577c21fb808MD51TEXTZamaMLS_DISSERT.pdf.txtZamaMLS_DISSERT.pdf.txtExtracted texttext/plain143037https://repositorio.ufrn.br/bitstream/123456789/13454/6/ZamaMLS_DISSERT.pdf.txtca6c94c0aece75c38d810a1b3899a70eMD56CaracterizacaoMolecularLaboratorial_Silveira_2010.pdf.txtCaracterizacaoMolecularLaboratorial_Silveira_2010.pdf.txtExtracted texttext/plain142985https://repositorio.ufrn.br/bitstream/123456789/13454/8/CaracterizacaoMolecularLaboratorial_Silveira_2010.pdf.txt48ac234cac8cfb1bdb9cea99c4c97eb2MD58THUMBNAILZamaMLS_DISSERT.pdf.jpgZamaMLS_DISSERT.pdf.jpgIM Thumbnailimage/jpeg2904https://repositorio.ufrn.br/bitstream/123456789/13454/7/ZamaMLS_DISSERT.pdf.jpgb2b16a9461cdfdc30076c7787db9a69eMD57CaracterizacaoMolecularLaboratorial_Silveira_2010.pdf.jpgCaracterizacaoMolecularLaboratorial_Silveira_2010.pdf.jpgGenerated Thumbnailimage/jpeg1274https://repositorio.ufrn.br/bitstream/123456789/13454/9/CaracterizacaoMolecularLaboratorial_Silveira_2010.pdf.jpg9ae166dafe8f12c099e2aa083e4b6126MD59123456789/134542019-05-26 02:22:44.192oai:https://repositorio.ufrn.br:123456789/13454Repositório de PublicaçõesPUBhttp://repositorio.ufrn.br/oai/opendoar:2019-05-26T05:22:44Repositório Institucional da UFRN - Universidade Federal do Rio Grande do Norte (UFRN)false |
dc.title.por.fl_str_mv |
Caracterização molecular e laboratorial da talassemia beta e da interação hemoglobina s/talassemia beta |
title |
Caracterização molecular e laboratorial da talassemia beta e da interação hemoglobina s/talassemia beta |
spellingShingle |
Caracterização molecular e laboratorial da talassemia beta e da interação hemoglobina s/talassemia beta Silveira, Zama Messala Luna da Talassemia beta Hemoglobinopatias Interação hemoglobina S/talassemia beta Mutações PCR/RFLP β -thalassemia Hemoglobinopathies S-β thalassemia Mutations PCR/RFLP CNPQ::CIENCIAS DA SAUDE::FARMACIA |
title_short |
Caracterização molecular e laboratorial da talassemia beta e da interação hemoglobina s/talassemia beta |
title_full |
Caracterização molecular e laboratorial da talassemia beta e da interação hemoglobina s/talassemia beta |
title_fullStr |
Caracterização molecular e laboratorial da talassemia beta e da interação hemoglobina s/talassemia beta |
title_full_unstemmed |
Caracterização molecular e laboratorial da talassemia beta e da interação hemoglobina s/talassemia beta |
title_sort |
Caracterização molecular e laboratorial da talassemia beta e da interação hemoglobina s/talassemia beta |
author |
Silveira, Zama Messala Luna da |
author_facet |
Silveira, Zama Messala Luna da |
author_role |
author |
dc.contributor.authorID.por.fl_str_mv |
|
dc.contributor.authorLattes.por.fl_str_mv |
http://lattes.cnpq.br/9321913536357216 |
dc.contributor.advisorID.por.fl_str_mv |
|
dc.contributor.advisorLattes.por.fl_str_mv |
http://lattes.cnpq.br/9869053526045053 |
dc.contributor.referees1.pt_BR.fl_str_mv |
Schimieguel, Dulce Marta |
dc.contributor.referees1ID.por.fl_str_mv |
|
dc.contributor.referees1Lattes.por.fl_str_mv |
http://lattes.cnpq.br/6361482547166270 |
dc.contributor.referees2.pt_BR.fl_str_mv |
Araújo, Aderson da Silva |
dc.contributor.referees2ID.por.fl_str_mv |
|
dc.contributor.referees2Lattes.por.fl_str_mv |
http://lattes.cnpq.br/9615780257704881 |
dc.contributor.author.fl_str_mv |
Silveira, Zama Messala Luna da |
dc.contributor.advisor1.fl_str_mv |
Medeiros, Tereza Maria Dantas de |
contributor_str_mv |
Medeiros, Tereza Maria Dantas de |
dc.subject.por.fl_str_mv |
Talassemia beta Hemoglobinopatias Interação hemoglobina S/talassemia beta Mutações PCR/RFLP |
topic |
Talassemia beta Hemoglobinopatias Interação hemoglobina S/talassemia beta Mutações PCR/RFLP β -thalassemia Hemoglobinopathies S-β thalassemia Mutations PCR/RFLP CNPQ::CIENCIAS DA SAUDE::FARMACIA |
dc.subject.eng.fl_str_mv |
β -thalassemia Hemoglobinopathies S-β thalassemia Mutations PCR/RFLP |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS DA SAUDE::FARMACIA |
description |
Beta thalassemia arises as a consequence of the reduction (β+, β++, βsilent) or absence (β0) of beta globin chain synthesis and results from a number of mechanisms that lead to genetic defects. The inheritance of beta thalassemia is characterized by the existence of heterozygous individuals, compound heterozygotes, homozygotes and those with coinheritance of beta thalassemia allele and other thalassemias and/or hemoglobin variants. The aim of this study was to perform molecular and laboratory characterization of beta thalassemia in heterozygous and homozygous individuals and in those with coinheritance of S beta thalassemia. A total of 48 individuals were included (35 heterozygotes, 4 homozygotes and 9 S beta thalessemia carriers) referred to the Integrated Laboratory of Clinical Analyses of the Federal University of Rio Grande do Norte (UFRN) and the Hematology Ambulatory Facility of the Dalton Barbosa Cunha Hemocenter (Hemonorte Natal, Brazil). Peripheral blood samples form each patient underwent the following laboratory examinations: erythrogram, hemoglobin electrophoresis at alkaline pH, measurements of Hb A2, Fetal Hb and serum ferritin. DNA was extracted using the illustra blood genomicPrep Mini Spin Kit and molecular characterization was performed by the PCR/RFLP technique, which involves digestion with specific restriction enzymes for IVS-1 nt 1 (G®A), IVS-1 nt 6 (T®C) and codon 39 (CAG®TAG) mutations. Of the 35 heterozygotes, 37.1% showed IVS-1 nt 6 mutation, 42.9% IVS-1 nt 1 and 20% were carriers of other mutations not identified by the technique used. The four homozygous patients presented with the IVS-1 nt 6 mutation, while 66.7% of the individuals with S beta thalassemia had the IVS-1 nt 1 mutation. Codon 39 was not detected in any of the patients investigated. Of the thallasemic alleles found, 40.4% were IVS- 1 nt 1, 40.4% IVS-1 nt 6 and 19.2% were not identified. Laboratory data showed that the heterozygotes exhibited microcytosis and hypochromia, evidenced by MCV ranging from 57 to 75fL and MCH from 15.9 to 23.6 pg. Hemoglobin A2 varied between 3.7 and 7.2%. The homogygotes also showed reduced MCV and MCH and elevated HbA2.. Comparison of laboratory data between heterozygous individuals with IVS-1 nt 1 and IVS-1 nt 6 mutations showed that heterozygotes for the IVS1-1 mutation had significantly lower mean MCV and MCH (p = 0.023 and 0.007, respectively) and significantly higher hemoglobin A2 (p < 0.001) when compared to heterozygotes for the IVS-1 nt 6 mutation. PCR/RFLP was useful in identifying the presence or absence of IVS-1 nt 6, IVS-1 nt 1 and codon 39 mutations in most of the patients investigated here. This is the first study conducted in the state of Rio Grande do Norte, Brazil aimed at identifying beta thalassemia mutations and represents an important contribution to the knowledge regarding the molecular profile of beta thalassemia in our country |
publishDate |
2010 |
dc.date.available.fl_str_mv |
2010-11-30 2014-12-17T14:16:25Z |
dc.date.issued.fl_str_mv |
2010-02-25 |
dc.date.accessioned.fl_str_mv |
2014-12-17T14:16:25Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
SILVEIRA, Zama Messala Luna da. Caracterização molecular e laboratorial da talassemia beta e da interação hemoglobina s/talassemia beta. 2010. 82 f. Dissertação (Mestrado em Bioanálises e Medicamentos) - Universidade Federal do Rio Grande do Norte, Natal, 2010. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufrn.br/jspui/handle/123456789/13454 |
identifier_str_mv |
SILVEIRA, Zama Messala Luna da. Caracterização molecular e laboratorial da talassemia beta e da interação hemoglobina s/talassemia beta. 2010. 82 f. Dissertação (Mestrado em Bioanálises e Medicamentos) - Universidade Federal do Rio Grande do Norte, Natal, 2010. |
url |
https://repositorio.ufrn.br/jspui/handle/123456789/13454 |
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por |
language |
por |
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info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
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Universidade Federal do Rio Grande do Norte |
dc.publisher.program.fl_str_mv |
Programa de Pós-Graduação em Ciências Farmacêuticas |
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UFRN |
dc.publisher.country.fl_str_mv |
BR |
dc.publisher.department.fl_str_mv |
Bioanálises e Medicamentos |
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Universidade Federal do Rio Grande do Norte |
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reponame:Repositório Institucional da UFRN instname:Universidade Federal do Rio Grande do Norte (UFRN) instacron:UFRN |
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Universidade Federal do Rio Grande do Norte (UFRN) |
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UFRN |
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UFRN |
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Repositório Institucional da UFRN |
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