Mendoza SI, Olarte CI, Rivera DTS, Cerón MR, De la Cruz RA, Collazo JJ, Martínez MC, Martínez TA

Language: Spanish
References: 24
Page: 11-17
PDF size: 492.19 Kb.

ABSTRACT

Background: Hemophilias are recessive genopathies linked to the X chromosome that occur due to deficiency in the production of coagulation factors. The objective of this study was to determine the coagulation factor 8 intron 22 inversion (Inv22) in two families with hemophilia A and B and its clinical impact.
Materials and Methods: Observational, descriptive study that evaluated the molecular alteration in intron 22 inversion in the gene that codes for coagulation factor VIII in three families with hemophilia A and B. Healthy individuals were negative controls. Peripheral blood samples were taken from each member of the family as well as from the patient, leukocytes were isolated from which DNA was extracted. In situ- PCR was performed by restriction with the enzyme BclII, the digested fragments were ligated with T4 ligase and finally inverse PCR was performed with specific primers for types 1 and 2 of intron 22 inversion.
Results: In family 1, the patient had intron 22 inversion type 2, the mother was positive and a carrier of Inv22 type 2; one the two daughters of the patient was negative for Inv22 and the other was positive for Inv22 with type 2. In family 2, the patient had Inv22 type 2, the patient’s sister, mother and grandmother were carriers of Inv22 type 2, two aunts and a cousin were negative for Inv22. In family 3 the patient had moderate hemophilia B, two sisters and the mother were negative for Inv22. Healthy individuals were negative for intron 22 inversion.
Conclusions: Inv22 type 2 was the most frequent in patients with severe hemophilia A. The in situ-PCR technique for the study of Inv22 is a useful and reliable tool in the analysis of patients and families affected by severe hemophilia A, since it allows the characterization of the type of intron 22 inversion (type 1 or 2) of the patient in a single blood sample.

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