Detection of the schistosome circulating cathodic antigen by enzyme immunoassay using biotinylated monoclonal antibodies.
We have developed an enzyme immunoassay (ELISA) for the quantification of the schistosome circulating cathodic antigen (CCA), a glycoprotein associated with the syncitium lining the gut of the parasite. A mouse monoclonal antibody of IgG3 isotype was used as coating (antigen-capture) antibody, while a biotinylated mouse monoclonal IgM was used as second (antigen-detecting) antibody. Streptavidin-alkaline phosphatase was used as enzyme label. The lower detection limit of the assay was 1.0 ng of the trichloroacetic acid soluble fraction of adult worm antigen (AWA-TCA) per ml, which corresponds to approximately 0.2 ng CCA per ml. The ELISA showed a linear range from 1.0 to 62.5 ng AWA-TCA per ml. Serum and urine samples of 16 individuals infected with Schistosoma mansoni (egg counts ranging from 5 to 4820 eggs per gram of faeces) were tested in the assay. Antigen titres ranged from less than 4-8192. This assay represents a considerable advantage in diagnosis of Schistosoma infections as it allows the detection and quantification of CCA in serum and urine in even lightly infected individuals.
de Jonge
N
N
Laboratory of Parasitology, Medical Faculty, University of Leiden, The Netherlands.
Kremsner
P G
PG
Krijger
F W
FW
Schommer
G
G
Fillié
Y E
YE
Kornelis
D
D
van Zeyl
R J
RJ
van Dam
G J
GJ
Feldmeier
H
H
Deelder
A M
AM
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000947:Antigens, Helminth / Q000032:analysis*; D001710:Biotin; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006023:Glycoproteins / Q000032:analysis*; D015801:Helminth Proteins / Q000032:analysis*; D006801:Humans; D012550:Schistosoma mansoni / Q000276:immunology*; D012555:Schistosomiasis mansoni / Q000175:diagnosis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth; C091532:CCA protein, Schistosoma mansoni; D006023:Glycoproteins; D015801:Helminth Proteins; D001710:Biotin