title: Single nucleotide polymorphism typing with a surface plasmon resonance-based sensor using hybridization enhancement blockers.
abstract: We constructed a simple method for discrimination of single nucleotide polymorphism (SNP) with a surface plasmon resonance (SPR)-based sensor by determining the binding volume (BV) between a target SNP allele and a probe complementary to the target. In the method, a novel additive termed "blocker," which is a short single-stranded DNA complementary to the target, was used. The blocker enhanced the BV of the target only to the full-match probe 10-fold or more, so the SNP alleles could be discriminated readily. The effect of the blocker concentration was also examined. The BV to only a full-match probe increased with increasing the blocker concentration and reached a plateau at the concentration of 300-500 nM. To assess the effectiveness of this method, the SNP associated with progressive rod-cone degeneration in dog was determined. The results of genotyping with the method were in good agreement with those obtained by direct sequencing.
authors.affiliation: Biotechnology and Food Research Institute, Fukuoka Industrial Technology Centre, 1465-5 Aikawa, Kurume, Fukuoka, 839-0861, Japan, sokumura@fitc.pref.fukuoka.jp.
authors.forename: Shiro
authors.initials: S
authors.lastname: Okumura
authors.forename: Rieko
authors.initials: R
authors.lastname: Kuroda
authors.forename: Kuniyo
authors.initials: K
authors.lastname: Inouye
mesh_terms: D000818:Animals; D004285:Dogs; D005838:Genotype; D009693:Nucleic Acid Hybridization; D020641:Polymorphism, Single Nucleotide; D020349:Surface Plasmon Resonance / Q000379:methods*