Development of novel microarray methodology for the study of mutations in the SERPINA1 and ADRB2 genes—Their association with Obstructive Pulmonary Disease and Disseminated Bronchiectasis in Greek patients ☆ Athanasios Papatheodorou a, ⁎ , Periklis Makrythanasis a,b , Marios Kaliakatsos a , Aikaterini Dimakou c , Dora Orfanidou d , Charis Roussos e , Emmanuel Kanavakis a , Maria Tzetis a a Department of Medical Genetics, Medical School, University of Athens, Thivon and Levadias, 11257 Athens, Greece b Department of Genetic Medicine and Development, Faculty of Medicine, University of Geneva, Switzerland c 6th Department of Respiratory Medicine, Sotiria Chest Disease Hospital, Greece d 3rd Department of Internal Medicine, Medical School, Athens University, Sotiria Chest Disease Hospital, Athens, Greece e Critical Care Department, Evangelismos Hospital, Medical School, Athens University, Greece Received 18 December 2008; received in revised form 3 July 2009; accepted 25 August 2009 Available online 10 September 2009 Abstract Objectives: The aim of our study was to determine the genetic risk conferred by SNPs in the SERPINA1 and ADRB2 for development of Chronic Obstructive Pulmonary Disease (COPD) and Disseminated Bronchiectasis (DB), while at the same time validating the NanoChip technology. This was a case–control study consisting of 112 COPD, 62 DB patients and 2 control groups (106 smokers without COPD: healthy smokers control group and 205 general population subjects). Design and methods: The novel methodology of the Nanogen NanoChip® 400 (NC400 Nanogen www.nanogen.com) was employed for genotyping five mutations/SNPs in the SERPINA1 and 2 in the ADRB2 gene. Results: For the SERPINA1 gene a statistically significant difference in the frequency was found for heterozygotes for p.V213A between DB patients and healthy smokers (44.1% vs. 26.4% respectively; p = 0.035) and for heterozygotes for c.1237GNA between DB patients and general population subjects (10.2% vs. 25.4% respectively; p = 0.023). There was a clustering of ADRB2 p.Gly16 homozygotes in patients with severe COPD (24/44, 54.5% with FEV 1 values b35% of predicted). Conclusions: The SERPINA1 p.V213A polymorphism was found associated with DB risk while the ADRB2 p.G16R is a risk factor for severe COPD in smokers. © 2009 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved. Keywords: SERPINA1 ADRB2; Chronic Obstructive Pulmonary Disease; Disseminated Bronchiectasis; haplotypes; SNPs Introduction Chronic Obstructive Pulmonary Disease (COPD) is a major cause of chronic morbidity and mortality. The World Health Organization (WHO) listed COPD as the fifth leading cause of death in the world and it is estimated that COPD will be the third commonest cause of death globally by 2020 [1]. In Greece, the prevalence of the disease ranges between 10.7% and 15.8% in subjects aged ≥ 50 years. COPD is a complex disease influenced by genetic and environmental factors. Cigarette smoking is the major environmental determinant of COPD; Available online at www.sciencedirect.com Clinical Biochemistry 43 (2010) 43 – 50 Abbreviations: SERPINA1, Serine Protease Inhibitor A1; ADRB2, β2- adrenoceptor; AHR, airway hyperresponsiveness; PCR, polymerase chain reaction; SNP, single nucleotide polymorphisms; COPD, Chronic Obstructive Pulmonary Disease; DB, Disseminated Bronchiectasis. ☆ Human Genes: Alpha-1 antitrypsin (AAT, SERPINA1; MIM# 107400) and β2-adrenergic receptor (ARB2; MIM# 109690). GenBank: www.ncbi.nlm.nih. gov/entrez/query.fcgi?db=nucleotide accession numbers NM_000295 and NM_000024 respectively for AAT and ADRB2. NCBI SNP Database: http:// www.ncbi.nlm.nih.gov/. ⁎ Corresponding author. Fax: +30 210 7795553. E-mail address: atpapath@med.uoa.gr (A. Papatheodorou). 0009-9120/$ - see front matter © 2009 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved. doi:10.1016/j.clinbiochem.2009.08.026