55º Congresso Brasileiro de Genética Resumos do 55º Congresso Brasileiro de Genética • 30 de agosto a 02 de setembro de 2009 Centro de Convenções do Hotel Monte Real Resort • Águas de Lindóia • SP • Brasil www.sbg.org.br - ISBN 978-85-89109-06-2 320 Different methodological approaches to diagnose the Androgen Insensitivity Syndrome: case report in a public service of genetics Melo, COA1,3; Silva, DM1,2,3; Ribeiro CL1; Silva, GP 2; Silva, DC1; de Melo, AV1; Costa, EOA 1,3; da Silva, CC1,2; da Cruz, AD1,2,3 1 Universidade Católica de Goiás – Departamento de Biologia – Núcleo de Pesquisas Replicon, Goiânia, GO. LaGene – SuLeide, Laboratório de Citogenética Humana e Genética Molecular- Superintendência Leide das Neves Ferreira Secretaria Estadual de Saúde de Goiás, Goiânia, GO. 2 3 Pró-Reitoria de Pós-Graduação e Pesquisa, Mestrado em Genética, Universidade Católica de Goiás. [email protected] Keywords: Androgen Receptor, Androgen Insensitivity Syndrome, infertility, FISH, PCR. The disorders that affect the genetic sex can cause many types of problems to individuals and their families. The Androgens Insensitivity Syndrome (AIS) is a disorder linked to chromosome X and caused by mutations in the gene of Androgen Receptor (AR). Mutations that severely affect the quantity, structure and function of AR cause CAIS (Complete Androgens Insensitivity Syndrome) phenotype, also known as Testicular Feminization Syndrome. All XY individuals with CAIS are infertile. In this context, in the year of 2008, the patient M. C. J., 15 years old, was referred to the Laboratory of Human Cytogenetics and Molecular Genetics of the city of Goiânia showing poorly developed breasts and primary amenorrhea. Her medical history showed that when she was a newborn, some doctors removed an abdominal mass, which was diagnosed as an umbilical hernia. Thus, the G-band karyotyping of this patient showed the result 46, XY. It was concluded that the mass withdrawal from the abdomen of the patient was, in fact, testes and that the patient had a condition known as cryptorchidism, a reproductive change characterized by a failure of the displacement of one or both testes to the abdominal cavity to the scrotum. We performed the methods of PCR and FISH to verify mutations of the exons 1, 4, 6, 7 and 8 of the AR gene and to detect the AR gene, respectively. We prepared a culture of T-lymphocytes in the medium RPMI 1640, supplemented with 20% of bovine fetal calf and 2% of phytohemagglutinin. Metaphasic preparations were made by conventional methodology. The use of slides for FISH were prepared with a micropipette, dripped about 15 µL of the material set. Only the slides of good quality (in terms of metaphases), were selected by phase contrast microscope and were subjected to the method of FISH using the probe LSI Androgen Receptor SpectrumOrange (Xq12) (Vysis ®, Abbott Park, Illinois, USA). For PCR, primers were used for the exons 1, 4, 6, 7 and 8 of AR. By PCR, it was possible to observe that the patient did not present the exons 1, 4 and 7 of AR, but despite of that, the probe hybridized the AR region. This could be explained by the size of the probe (380 kb), which was higher than the AR gene (90 kb), indicating that the deletions of some exons did not influence the hybridization of the probe. So, these results showed that the probe used in our lab, to diagnose AIS, has to be associated to PCR to detect mutations (deletions) of some regions in the AR gene, being the appropriate tools to diagnose cases of AIS.