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Discriminação alélica do fator V da coagulação por PCR em tempo real: diagnóstico simples e preciso
(2009-03) OLIVEIRA FILHO, Aldemir Branco de; CAMPOS, Júlia Furtado; LIMA, Magaly do Bom Parto Lopes Vieira; MELO, Fárida Coeli de Barros Correia; NEVES, Washington Batista das; MELO, Raul Antônio Morais; LEMOS, José Alexandre Rodrigues de
Among cardiovascular diseases, venous thrombosis is important due to the association between acquired and genetic risks factors. Hereditary resistance to activated protein C has been identified as the main cause of venous thrombosis, and is frequently associated to the factor V Leiden mutation (G1694A). In homozygotic individuals, the risk of venous thrombosis is 50 to 100 times higher that in normal patients, while in heterozygotic patients the risk is 5 to 10 times higher. Based on the need of evaluation and follow up of patients with venous thrombosis and prevention in their respective families, a simple method of allelic discrimination of coagulation V factor was developed using real time PCR. Sixty-seven patients with a history of venous thrombosis and 51 individuals without venous thrombosis were selected for this study. First, identification of the factor V allele was achieved through conventional PCR followed by enzymatic digestion (Mnl). Subsequently, diagnosis was attained by real time PCR. Both the methods investigated the G1691A polymorphism using VIC and FAM fluorophores to mark nucleotides G and A, respectively. By PCR-RFLP, 95 individuals were diagnosed as normal homozygotes, 21 as heterozygotes and 2 as homozygotic factor V Leiden individuals. The same results were obtained using real time PCR. Maximum similarity between the results of real time PCR and PCR-RFLP indicates high precision of the new method for allelic identification and visualization of factor V Leiden.
Acesso aberto (Open Access)
Increased risk of venous thrombosis by AB alleles of the ABO blood group and Factor V Leiden in a Brazilian population
(2009) LIMA, Magaly do Bom Parto Lopes Vieira; OLIVEIRA FILHO, Aldemir Branco de; CAMPOS, Júlia Furtado; MELO, Fárida Coeli de Barros Correia; NEVES, Washington Batista das; MELO, Raul Antônio Morais; LEMOS, José Alexandre Rodrigues de
Most cases of a predisposition to venous thrombosis are caused by resistance to activated protein C, associated in 95% of cases with the Factor V Leiden allele (FVL or R506Q). Several recent studies report a further increased risk of thrombosis by an association between the AB alleles of the ABO blood group and Factor V Leiden. The present study investigated this association with deep vein thrombosis (DVT) in individuals treated at the Hemocentro de Pernambuco in northeastern Brazil. A case-control comparison showed a significant risk of thrombosis in the presence of Factor V Leiden (OR = 10.1), which was approximately doubled when the AB alleles of the ABO blood group were present as well (OR = 22.3). These results confirm that the increased risk of deep vein thrombosis in the combined presence of AB alleles and Factor V Leiden is also applicable to the Brazilian population suggesting that ABO blood group typing should be routinely added to FVL in studies involving thrombosis.