New application of indirect fluorescent antibody (IFA) technique using latex particles coupled with verotoxin 2 from Escherichia coli O157:H7 in order to determine colostral antibody titers in immunized dairy cows.
A simple and novel assay method for determining colostral and serum against soluble verotxin 2 (VT2) titers by indirect fluorescent antibody (IFA) assay using latex sensitized with VT2 was devised. The latex particles did not auto-fluoresce, and non specific reactions disappeared after washing with phosphate buffered saline containing 3 M Nacl. The highest titer measured by neutralizing test was observed at 1 day after delivery. The highest titer for each immunoglobulin class measured by enzyme-linked immunosorbent assay (ELISA) or IFA using latex sensitized with VT2 was also observed at 1 day after delivery. The changes in titer measured by each method showed similar patterns. Furthermore, the titers for IgG antibody were higher than those for IgM or IgA antibodies. Thus, the titers of bovine immune colostral antibody and each immunoglobulin class could be measured by IFA using latex sensitized with VT2.
a Laboratory of Immunology and Graduate School of Environmental Health Science , Azabu University , Chuo-ku, Sagamihara , Kanagawa , Japan.
Tetsurou
T
Seita
Takashi
T
Kuribayashi
Seiji
S
Yamaguchi
Katsunori
K
Furuhata
Shizuo
S
Yamamoto
D000818:Animals; D000907:Antibodies, Bacterial / Q000032:analysis*; D002417:Cattle; D003126:Colostrum / Q000737:chemistry / Q000276:immunology* / Q000382:microbiology; D003612:Dairying; D004797:Enzyme-Linked Immunosorbent Assay; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000276:immunology; D019453:Escherichia coli O157 / Q000737:chemistry / Q000276:immunology; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect / Q000379:methods*; D007114:Immunization; D007070:Immunoglobulin A / Q000032:analysis; D007074:Immunoglobulin G / Q000032:analysis; D007075:Immunoglobulin M / Q000032:analysis; D008863:Microspheres; D009500:Neutralization Tests; D011247:Pregnancy; D022641:Shiga Toxin 2 / Q000737:chemistry* / Q000276:immunology
D000907:Antibodies, Bacterial; D004800:Enzymes, Immobilized; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D022641:Shiga Toxin 2