Assay for measuring relative potency of leptospiral bacterins containing serovar pomona.
An enzyme-linked immunosorbent assay (ELISA) was developed for the quantitation of leptospiral antigen in bacterins containing Leptospira interrogans serovar pomona type kennewicki. A monoclonal antibody (MAb), 2D7, which is directed against a surface antigen on whole cells of L. interrogans serovar pomona type kennewicki, was used in the assay. The capture of antigen in bacterins by a polyclonal antiserum was followed by the addition of the 2D7 ascites fluid, an anti-mouse conjugate and substrate. Biologicals evaluated with this system included preparations containing type kennewicki antigen (homologous) and those not containing type kennewicki antigen (heterologous). Heterologous bacterins gave optical density (OD) values comparable to those of blank wells. Homologous bacterins yielded OD values equal to or greater than those of the National Veterinary Services Laboratories (NVSL) reference pomona bacterin. The relative potencies (RP) of 84 licensed commercial Leptospira pomona bacterin serials were evaluated against the NVSL reference pomona bacterin using the NVSL Relative Potency computer program. Random samples of 1, 2, 3 and 5 ml dose products were selected for evaluation with this system. All products tested passed the hamster potency assay required for leptospiral bacterins. This ELISA system enables detection of antigen in bacterins containing L. interrogans serovar pomona type kennewicki and demonstrates the potential for in vitro testing of leptospiral bacterins.
Ruby
K W
KW
National Veterinary Services Laboratories, U.S. Department of Agriculture, Ames, Iowa 50010.
Cardella
M A
MA
Knudtson
W U
WU
D000818:Animals; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial / Q000032:analysis; D001428:Bacterial Vaccines / Q000032:analysis* / Q000592:standards; D001681:Biological Assay / Q000379:methods*; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005069:Evaluation Studies as Topic; D007921:Leptospira interrogans / Q000145:classification / Q000276:immunology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D012015:Reference Standards; D012703:Serotyping; D014895:Weil Disease / Q000517:prevention & control
D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D001428:Bacterial Vaccines