detection of haplotype mutations of the md-2 gene promoter associated with der p2-induced allergy using a nanostructured biosensor

Yu-Ting Chin,1 En-Chih Liao,2 Chia-Che Wu,1 Gou-Jen Wang,1,3,4 Jaw-Ji Tsai2 1Department of Mechanical Engineering, National Chung-Hsing University, 2Department of Medical Education and Research, Taichung Veterans General Hospital, 3Graduate Institute of Biomedical Engineering, 4PhD Program in Tissue Engineering and Regenerative Medicine, National Chung-Hsing University, Taichung, Taiwan Abstract: Group 2 allergens (Der p2) have been reported to be a major cause of the human immune response to dust mite allergens. In this study, we have demonstrated for the first time the effective differentiation between haplotype mutation and normal genes in the MD-2 gene promoter using a nanostructured biosensor. A 70-mer gene fragment containing the haplotype of two single nucleotide polymorphisms in the MD-2 gene promoter region was used as a probe to detect haplotype mutations associated with Der p2-induced allergy. Discrimination was achieved using electrochemical impedance spectroscopy. The discrimination experiments employed 30 haplotype mutation samples and 30 normal target samples. The haplotype mutation samples and normal target samples could be clearly discriminated, even using samples produced by a five-cycle polymerase chain reaction process. The time and cost of sample preparation for the polymerase chain reaction process in the clinical setting can thus be reduced. Keywords: haplotype mutation detection, nanostructured biosensor, MD-2 gene promoter, electrochemical impedance spectroscopy