β- 2 adrenergic receptor (B2AR) agonists are the most widely prescribed rescue agents used in the treatment of asthma. Recent studies have indicated a relationship between a polymorphism at codon 16 of the B2AR gene, and the response to recurrent β-agonist therapy. The B2AR polymorphism of interest involves a single nucleotide change from A to G, resulting in an amino acid change from Arginine (Arg) to Glycine (Gly). Clinical efforts to further investigate this relationship require an accurate, reliable and inexpensive method for detecting the polymorphism. In this study, we report an LCx® assay for the detection of a single nucleotide polymorphism at codon 16 of the β- 2 adrenergic receptor. This assay is capable of detecting patients harboring any of the three possible genotypes at this locus, namely, homozygous wild type, homozygous variant or heterozygous individuals with a single genomic DNA sample of 25–500 ng. It requires minimum hands-on time with automated detection. The assay would be suitable for use in research labs for screening of a large number of samples. We believe that this type of assay will facilitate research and clinical investigations in elucidating the association of SNPs with disease states, diagnosis, prognosis and treatment.