Rolling circle amplification for signal enhancement in ovalbumin detection.
We have developed a rolling circle amplification-procedure as a method to enhance signals of immunoassay (immuno-RCA) to detect one of the major food allergens, ovalbumin (OVA). Immuno-RCA in combination with a fluorescent dye and a circular DNA probe prepared by intramolecular ligation easily allowed the real-time detection of OVA, and could apparently detect signals from OVA with concentrations ranging from 10(-12) to 10(-7) g/mL. Therefore, sensitive and easily handled aspects of this method would contribute to an effective signal enhancement in immunoassay for food allergen detections.
Kobori
Toshiro
T
Nanobiotechnology Laboratory, National Food Research Institute, National Agriculture and Food Research Organization, 2-1-12 Kan-nondai, Tsukuba 305-9642, Japan. tkobo@affrc.go.jp
Matsumoto
Atsuko
A
Takahashi
Hirokazu
H
Sugiyama
Shigeru
S
D000818:Animals; D001483:Base Sequence; D015342:DNA Probes / Q000235:genetics; D004270:DNA, Circular / Q000235:genetics; D005456:Fluorescent Dyes / Q000378:metabolism; D005512:Food Hypersensitivity; D007118:Immunoassay / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009838:Oligodeoxyribonucleotides / Q000235:genetics; D010047:Ovalbumin / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D012815:Signal Processing, Computer-Assisted; D013997:Time Factors
D015342:DNA Probes; D004270:DNA, Circular; D005456:Fluorescent Dyes; D009838:Oligodeoxyribonucleotides; D010047:Ovalbumin