Avaliação do transplante de medula óssea alogênico por meio do estudo de regiões de repetições seqüenciais no genoma humano (VNTRs e STRs)
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Data
2004-01-01
Autores
Santana, Rita K. [UNESP]
Mattos, Luiz C.
Ruiz, Milton A.
Moreira, Haroldo Wilson [UNESP]
Título da Revista
ISSN da Revista
Título de Volume
Editor
Associação Brasileira de Hematologia e Hemoterapia e daSociedade Brasileira de Transplante de Medula Óssea
Resumo
Os objetivos deste estudo foram estabelecer um protocolo para a análise de minissatélites ou VNTRs e microssatélites ou STRs em pacientes que se submeteram ao TMO alogênico; verificar a validade da metodologia e dos loci estudados e avaliar o tipo de recuperação do paciente. Foram analisados o DNA do paciente anterior e posterior ao transplante de 14 indivíduos e dos respectivos doadores. Amplificações por PCR de seis loci: D1S80, SE33, HumTH01, 33.6, HumARA e HumTPO foram realizadas. Os produtos amplificados foram separados por eletroforese vertical em gel de poliacrilamida, e os fragmentos visualizados por coloração pela prata. Esse procedimento mostrou ser válido na verificação da recuperação alogênica, autóloga e provavelmente na quimérica. da somatória dos loci estudados, 63,1% apresentaram resultados possíveis de serem avaliados e, desses, 19,0% mostraram resultado informativo, 13,1% parcialmente informativo e 31,0% não informativo. Os 36,9% restantes não foram possíveis de avaliação. Dos loci avaliados, o que demostrou maior índice de resultado informativo foi o SE33, parcialmente informativo o HumTPO e não informativo o HumTH01, sendo o locus 33.6 o que mais apresentou resultados não possíveis de serem avaliados. Por outro lado, determinou-se a recuperação do paciente posterior ao transplante em 71,4% dos indivíduos, sendo que, desses, 90% apresentaram recuperação alogênica e 10% recuperação autóloga.
The purposes of this study were to establish a protocol for the analysis of minisatellites or VNTRs and microsatellites or STRs in patients who undergo allogeneic bone marrow transplantation; to verify the validity of the methodology and of the studied loci and to verify the type of recovery of the patient. The pre-transplant and post-transplant DNA of 14 recipients and their respective donors were analyzed. Six loci were amplified by PCR (D1S80, SE33, HumTH01, 33.6, HumARA and HumTPO). The amplified products were separated using a vertical polyacrylamide gel electrophoretic system and the resulting fragments were analyzed using silver staining. This procedure proved to be efficient in verifying autologous and allogeneic regeneration and probably chimerism regeneration too. of the loci studied, 63.1% presented results which could be analyzed, 19% were informative results, 13.1% partially informative and 31.0% non-informative. The locus that most demonstrated the informative results was the SE33 locus, the one that showed the partially informative results was the HumTPO and the non-informative results was the HumTH01. The 33.6 locus was the one that presented the majority of the results that could not be tested. Furthermore, it was possible to determine the patient regeneration after transplantation in 71.4% of the cases, 90% showed allogeneic regeneration and 10% autologous regeneration.
The purposes of this study were to establish a protocol for the analysis of minisatellites or VNTRs and microsatellites or STRs in patients who undergo allogeneic bone marrow transplantation; to verify the validity of the methodology and of the studied loci and to verify the type of recovery of the patient. The pre-transplant and post-transplant DNA of 14 recipients and their respective donors were analyzed. Six loci were amplified by PCR (D1S80, SE33, HumTH01, 33.6, HumARA and HumTPO). The amplified products were separated using a vertical polyacrylamide gel electrophoretic system and the resulting fragments were analyzed using silver staining. This procedure proved to be efficient in verifying autologous and allogeneic regeneration and probably chimerism regeneration too. of the loci studied, 63.1% presented results which could be analyzed, 19% were informative results, 13.1% partially informative and 31.0% non-informative. The locus that most demonstrated the informative results was the SE33 locus, the one that showed the partially informative results was the HumTPO and the non-informative results was the HumTH01. The 33.6 locus was the one that presented the majority of the results that could not be tested. Furthermore, it was possible to determine the patient regeneration after transplantation in 71.4% of the cases, 90% showed allogeneic regeneration and 10% autologous regeneration.
Descrição
Palavras-chave
VNTRs, STRs, transplante de medula óssea, PCR, VNTRs, STRs, bone marrow transplantation, PCR
Como citar
Revista Brasileira de Hematologia e Hemoterapia. Associação Brasileira de Hematologia e Hemoterapia e daSociedade Brasileira de Transplante de Medula Óssea, v. 26, n. 2, p. 109-113, 2004.