Detection of Pseudomonas aeruginosa antigenemia in granulocytopenic rabbits by radiommunoassay.
We assessed the ability of a solid-phase radioimmunoassay, modified to allow antigen detection in serum, to detect circulating antigens in granulocytopenic rabbits with Pseudomonas aeruginosa bacteremia. In vitro experiments with Pseudomonas lipopolysaccharide indicated that treatment of serum-lipopolysaccharide mixtures with heat, chloroform, or heparin improved the sensitivity for detecting lipopolysaccharide 8- to 16-fold. Chloroform treatment permitted antigen detection in serum or plasma of bacteremic rabbits in which antigen could be detected poorly or not at all in untreated specimens. However, chloroform-treated specimens occasionally caused dissolution of plastic tubes, resulting in nonspecific binding of 125I-labeled anti-Pseudomonas immunoglobulin G. Heating sera at 56 degree C for 30 min improved antigen detection both in both in lipopolysaccharide-serum mixtures and in bacteremic rabbits. Antigen was detected in the heated serum or plasma of 20% of 20 culture-positive granulocytopenic rabbits, none of 15 culture-negative granulocytopenic rabbits, and none of 38 normal rabbits. Antigen was detected in none of 15 rabbits with 2 to 300 colony-forming units of P. aeruginosa per ml of blood and in 4 of 5 rabbits with > 10(3) colony-forming units per ml. We conclude that circulating antigens are present in the blood of rabbits with high-level P. aeruginosa bacteremia and that these antigens can be detected by solid-phase radioimmunoassay. Further improvements in assay sensitivity will be required to detect antigens, if present, in animals with lower levels of P. aeruginosa bacteremia.
D000380:Agranulocytosis / Q000276:immunology*; D000818:Animals; D000942:Antigens, Bacterial / Q000032:analysis*; D004195:Disease Models, Animal; D006358:Hot Temperature; D011135:Polysaccharides, Bacterial / Q000032:analysis; D011552:Pseudomonas Infections / Q000276:immunology*; D011550:Pseudomonas aeruginosa / Q000276:immunology*; D011817:Rabbits; D011863:Radioimmunoassay / Q000379:methods; D018805:Sepsis / Q000276:immunology
D000942:Antigens, Bacterial; D011135:Polysaccharides, Bacterial