Detection of Cryptosporidium parvum oocysts in bovine feces by monoclonal antibody capture enzyme-linked immunosorbent assay

A monoclonal antibody enzyme-linked immunosorbent assay (ELISA) was developed to detect Cryptosporidium parvum oocysts in bovine feces. Fecal oocysts were concentrated by centrifugation through Formalin-ethyl acetate solution and captured with monoclonal antibody 18.280.2 reactive with C. parvum ooc...

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Bibliographic Details
Published in:Journal of Clinical Microbiology 1990-12, Vol.28 (12), p.2770-2774
Main Authors: Anusz, K.Z. (Agricultural Academy, Warsaw, Poland), Mason, P.H, Riggs, M.W, Perryman, L.E
Format: Article
Language:English
Subjects:
Animals
Antibodies, Monoclonal
ANTICORPS MONOCLONAL
ANTICUERPOS MONOCLONALES
Antigens, Protozoan - isolation & purification
BOVIN
Cattle
Cryptosporidiosis - diagnosis
CRYPTOSPORIDIUM
Cryptosporidium - immunology
Cryptosporidium - isolation & purification
DIAGNOSTIC
DIAGNOSTICO
DIARREA
DIARRHEE
ELISA
Enzyme-Linked Immunosorbent Assay - methods
ETAPAS DEL DESARROLLO ANIMAL
EXPERIMENTACION IN VIVO
EXPERIMENTATION IN VIVO
FECES
Feces - parasitology
GANADO BOVINO
HECES
INFECCION
INFECTION
STADE DE DEVELOPPEMENT ANIMAL
TERNERO
TEST ELISA
VEAU
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Description
Summary:A monoclonal antibody enzyme-linked immunosorbent assay (ELISA) was developed to detect Cryptosporidium parvum oocysts in bovine feces. Fecal oocysts were concentrated by centrifugation through Formalin-ethyl acetate solution and captured with monoclonal antibody 18.280.2 reactive with C. parvum oocysts. Captured oocysts were detected with goat anti-oocyst serum, following the addition of a peroxidase conjugate of rabbit anti-goat immunoglobulin and O-phenylenediamine substrate. The assay was specific for Cryptosporidium sp. oocysts and did not detect oocysts of Eimeria auburnensis, Eimeria bovis, Eimeria ellipsoidalis, or Eimeria zuernii. Assay sensitivity allowed detection of 3 X 10(5) oocysts per mi of feces, compared with 1 X 10(6) oocysts per mi detected by examination of acid-fast-stained fecal smears and 1 X 10(3) oocysts per ml detected by indirect immunofluorescence. The capture ELISA was suitable for diagnostic analysis of bovine fecal samples and for assessment of oocyst shedding in experimentally infected calves. This assay may also prove useful for diagnostic assessment of humans in which cryptosporidiosis is suspected
ISSN:0095-1137
1098-660X
DOI:10.1128/jcm.28.12.2770-2774.1990