Establishment of a sandwich enzyme linked immunosorbent assay for canine interleukin-8.
To estimate canine interleukin-8 (cIL-8) levels in blood plasma samples, a sandwich enzyme linked immunosorbent assay (ELISA) was established. For the development of the sandwich ELISA, polyclonal anti-cIL-8 (capturing), biotinylated anti-cIL-8 (developing) antibodies and glutathione-S-transferase/cIL-8 (GST/cIL-8) fusion protein as an antigen were used. cIL-8 in the fusion protein of GST/cIL-8 was detected in a dose dependent manner. The lowest limit of GST/cIL-8 detectable by this method was 2 ng/ml of GST/cIL-8 (containing; 0.470 ng/ml of cIL-8). IL-8 levels in the plasma samples from apparently healthy dogs were less than 0.470 ng/ ml. Higher levels of IL-8 were detected in the plasma samples of dogs with cystitis, dermatitis, and gastric cancer. These results suggest that the determination of cIL-8 by the sandwich ELISA is useful in diagnosis of inflammatory diseases in dogs.
Mohamed
A
A
Department of Molecular Immunology, Faculty of Agriculture, University of Tokyo, Japan.
Matsumoto
Y
Y
Yoshihara
K
K
Matsumoto
Y
Y
Watari
T
T
Tsujimoto
H
H
Hasegawa
A
A
Onodera
T
T
Hirota
Y
Y
D000818:Animals; D015415:Biomarkers / Q000097:blood; D003556:Cystitis / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary; D003872:Dermatitis / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary; D004283:Dog Diseases; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005982:Glutathione Transferase; D007202:Indicators and Reagents; D007249:Inflammation; D016209:Interleukin-8 / Q000032:analysis / Q000097:blood*; D008297:Male; D009919:Orchiectomy; D011993:Recombinant Fusion Proteins / Q000032:analysis; D012016:Reference Values; D051437:Renal Insufficiency / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary; D012680:Sensitivity and Specificity; D013274:Stomach Neoplasms / Q000097:blood / Q000175:diagnosis / Q000662:veterinary
D015415:Biomarkers; D007202:Indicators and Reagents; D016209:Interleukin-8; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase