Development of oligonucleotide lateral-flow immunoassay for multi-parameter detection.
We have developed a highly sensitive and rapid oligonucleotide lateral-flow immunoassay (OLFIA), using a colloidal gold as an indicator, for the simultaneous detection of antigens and/or antibodies in specimen. This system can detect more than two types of antigens and/or antibodies in a single assay device at the same time. The device is basically composed of colloidal gold-labeled antibodies and oligonucleotide-labeled antibodies fixed in a conjugate pad, and the complementary oligonucleotide-labeled proteins are immobilized on a nitrocellulose membrane. If the target antigen is present in a specimen, the colloidal gold-labeled antibody and oligonucleotide-labeled antibody will make a complex with the antigen. Subsequently, the formed complex migrates to the place where complementary oligonucleotide is immobilized and is bound to the solid phase via the DNA-DNA interaction. As a result, more than two types of reactions can be detected on a single assay device by the combination of colloidal gold-labeled antibodies, different oligonucleotide-labeled antibodies and complementary oligonucleotide-labeled proteins immobilized at different places on a nitrocellulose membrane.
Oku
Y
Y
Nissui Pharmaceutical Co., Ltd., Research Division, 1075-2 Hokunanmoro, Ibaraki, 307-0036, Yuki, Japan. y-oku@mail.nprl-unet.ocn.ne.jp
Kamiya
K
K
Kamiya
H
H
Shibahara
Y
Y
Ii
T
T
Uesaka
Y
Y
D000818:Animals; D000906:Antibodies / Q000032:analysis; D000911:Antibodies, Monoclonal; D000937:Antigen-Antibody Reactions; D000941:Antigens / Q000032:analysis; D001483:Base Sequence; D017979:Gold Colloid; D006801:Humans; D007118:Immunoassay / Q000379:methods* / Q000706:statistics & numerical data; D051379:Mice; D009838:Oligodeoxyribonucleotides / Q000235:genetics; D012680:Sensitivity and Specificity
D000906:Antibodies; D000911:Antibodies, Monoclonal; D000941:Antigens; D017979:Gold Colloid; D009838:Oligodeoxyribonucleotides