Romanian Biotechnological Letters Vol. 22, No. 6, 2016 Copyright © 2016 University of Bucharest Printed in Romania. All rights reserved ORIGINAL PAPER Romanian Biotechnological Letters, Vol. 22, No. 6, 2016 12026 A fully automated method for FTO and ADRB3 genotyping in syndromic (Prader-Willi Syndrome) and common obesity Received for publication, October 13 th , 2015 Accepted, February 29 th , 2016 ARSENE COSMIN 1,2 , URSU RADU 3 , VIOLETA DAN 2 , TRIFANESCU RALUCA 1 , BADIU CORIN 1 , NICULESCU MARIUS 4,5 , PUIU MARIA 6 , LYGIA ALEXANDRESCU 7 AND CUCU NATALIA 2,8 * 1 “Carol Davila” University of Medicine and Pharmacy, National Institute of Endocrinology “CI Parhon”, Department of Endocrynology, Bucharest, Romania 2 Association for Epigenetics and Metabolomics, Bucharest, Romania 3 “Carol Davila” University of Medicine and Pharmacy, National Institute of Endocrinology “CI Parhon”, Department of Medical Genetics, Bucharest, Romania 4 Titu Maiorescu University, Faculty of Medicine, Bucharest, Romania 5 Colentina Clinical Hospital, Bucharest, Romania 6 “Victor Babes” University of Medicine and Pharmacy, Deparment of Genetics, Timisoara, Romania 7 Association for Education in Nutrition, Bucharest, Romania 8 University of Bucharest, Faculty of Biology, Department of Genetics, Bucharest, Romania *Address correspondence to: University of Bucharest, Faculty of Biology, Department of Genetics, 1-3 Aleea Portocalilor, sector 5, Bucharest, 060101, Romania Tel.: +4021 318 15 65; e-mail: nataliacucu@gmail.com Abstract FTO (fat mass and obesity associated gene) and ADRB3 (adrenal-receptor beta 3) genes are considered important candidates as obesity risk genes due to their association with the alteration of the energy balance. This paper describes the implementation of FTO and ADRB3 genotyping, using the fully automated system, based on the quenching probes high resolution melting analysis (HRMA). Detection of single nucleotide polymorphisms (SNPs) in FTO and ADRB3 are useful to guide different treatment decisions in obese patients, depending both on the presence of SNRPN altered imprinting and the mutant FTO and ADRB3 genotypes. Blood and saliva samples wereobtained from selected individuals and the DNA was extracted and prepared for Prader-Willi syndrome (PWS) molecular confirmation using MS-PCR for the promotor/exon1 SNRPN region; also DNA extracted during a previous research program, was selected for further genetic tests of FTO (rs9939609) and ADRB3 (rs4994). In total, samples from 61 individuals were studied, from which 36 had the common obesity diagnosis, and FTO was strongly associated (cca 88%), 4 had the molecular PWS confirmation,and FTO mutant was moderately associated (cca 50%), and 7 had negative molecular PWS test result, but mutant FTO was relatively highly associated (cca 70%); 20 control, nonobese individuals DNA samples were moderately associated also with the mutant FTO (cca 40%). ADRB3 mutant C allele incidence in the obese subjects was only 25%, in PWS patients it was 0% and in the case of clinically suspected patients it was recorded at 14.3%. Key words: quenching probe method, obesity, Prader-Willi syndrome, single nucleotide polymorphisms 1. Introduction FTO (fat mass and obesity associated gene) and ADRB3 (adrenal-receptor beta 3) genes are intensively studied because they are associated with the alteration of the energy balance, and they are important candidates among the so-called obesity genes (T. RANKINEN & al. [1], T.O. KILPELAINEN & al. [2], K.A. FAWCETT & al. [3], L. QUAN & al. [4]). The A/T polymorphism (rs9939609) in the fat mass and obesity associated (FTO) gene (which