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Label-free detection of single-nucleotide polymorphisms associated with myeloid differentiation-2 using a nanostructured biosensor

By Yu-Ting Chin, En-Chih Liao, Chia-Che Wu, Gou-Jen Wang and Jaw-Ji Tsai


House dust mites are the major source of indoor allergens that are responsible for asthma. The major dust mite allergen is the group II allergen, Der p2. Myeloid differentiation-2 (MD-2) acts as an essential component in the CD14-TLR4 (toll-like receptor)/MD-2 receptor complex for Der p2 recognition. Mutations of the MD-2 associated gene on chromosome 8 degrade a human's innate responses. In this study, we report the effective detection of mutations to the MD-2 gene promoter, using a sensitive nanostructured biosensor with a sensing electrode of gold nanoparticles (GNPs) uniformly deposited in a nanohemisphere array. The 70 mer MD-2 expressed gene fragment was used to probe gene mutation. The complementary target, containing 156 mer nucleotide, was prepared using the fresh blood from patients with allergic disease. A total of 37 target samples, including 19 gene mutated samples and 18 normal samples, were prepared by a 20 cycles PCR process, and used for discrimination experiments. Experimental results illustrated that the nanostructured biosensor clearly discriminates between mutated and non-mutated MD-2 allergy genes

Topics: Myeloid differentiation-2 (MD-2) gene promoter, Single nucleotide polymorphism (SNP) detection, Nanostructured biosensor, Electrochemical impedance spectroscopy (EIS) analysis
Year: 2014
DOI identifier: 10.1016/j.bios.2013.05.049
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