155-P

Abstract

Aim Anorexia nervosa (AN) is a complex disorder with many genetic factors implicated in disease onset and pathology. In some cases the presence of a particular allele appears to modulate the immune response to certain exogenous antigens and protects the host from illness, whereas in other cases the host may mount an immune response against self-antigens. In the present study, we examined the possible association of DQB1∗ polymorphism with patients with Anorexia Nervosa, in Castilla y Leon (Spain) population. Methods We took samples of whole blood, 50-200mcl 28 AN,18 restrictive AN,: 10 compulsive AN, and 126 control subjects. Blood spots on 3MM filter paper were dried and stored at room temperature. DNA was extracted by two different procedures: salt precipitation and resin balls with metallic core (kit DNA Direct II from Dynal). We choosed the last one for its facility and rapidity, obtaining 1-4mcg of DNA. Classic SSP DQ “Low resolution” kit of Dynal was employed in this study. We have changed some general conditions of the DQ typing kit from the Dynal SSP method. We have modified the amount of DNA per tube to 2-4 ng and the concentration of the Taq Polymerase to 0,1 U/tube, instead of the 100 ng of DNA and 0,4 U/tube of Taq Polymerase recommended by the kit instructions. The conditions of the PCR had been: 1) 94 °C, 2 min, 1 cycle. 2) 94 °C 10 s, 65 °C 60 s 10 cycles. 3) 94 °C 10 s, 61 °C 50 s, 72 °C 30 s 30 cycles. 4) 72 °C 3 min. Results The allelic distribution in our study are for HLA DQB1∗02, ∗03, ∗04, ∗05, and ∗06 in control group (n = 126), 29,7; 27,3; 1,5; 19,4; 21,8. in AN group (n = 12) 34,3; 24,2; 0,7; 20,3; 20,6. Conclusions The Chi-square test show differences P