[Detection specificity of an optimal solid-phase enzyme immunoassay for Pseudomonas aeruginosa and Pseudomonas mallei].
The evaluation and application of an enzyme-immunoassay (EIA) for the detection of Pseudomonas (Ps.) aeruginosa and Ps. mallei is described. Polystyrene beads (1/4'') as the solid-phase are prepared by coating the balls with purified IgG from the serum of rabbits (9-12 micrograms/bead) in Coating-Buffer pH 9.6. After washing the balls they are saturated with 10% BSA or 10% FCS in PBS-Tween 20. The bacteria bound to the coated balls are detected by the specific peroxidase labelled IgG. This EIA using Ps. aeruginosa (P9) as a model is able to detect this bacterium within 5 hours, with stored coated balls 3.5 hours, with a detection limit of 10(4) CFU. Nine Pseudomonas-strains react stronger than other strains. These cross-reactions can be substantially reduced by absorbing the P9-conjugate with the cells of Ps. stutzeri (P15). With the other Pseudomonas-strains a high specificity is found with the P9-conjugate. After modifying this EIA for the detection of Ps. mallei (P18) the strains Ps. mallei (P57), Ps. pseudomallei (P17) and Ps. cepacia (P67) react with the P18-conjugate. With the other tested strains a high specificity is found at 10(7) CFU. The polystyrene bead-EIA is recommended as a sensitive and specific test for the detection of Ps. aeruginosa in about 5 resp. 3.5 hours. It only requires normal laboratory equipment and is thus a highly practicable method for routine diagnostic of Ps. aeruginosa.
Niederwöhrmeier
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B
Wehrwissenschaftliche Dienststelle der Bundeswehr für ABC-Schutz, Munster.
Böhm
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R
D003429:Cross Reactions; D007124:Immunoenzyme Techniques; D011237:Predictive Value of Tests; D011549:Pseudomonas / Q000302:isolation & purification*; D011552:Pseudomonas Infections / Q000175:diagnosis*; D011550:Pseudomonas aeruginosa / Q000302:isolation & purification*
Nachweisspezifität eines für Pseudomonas aeruginosa und Pseudomonas mallei optimierten Festphasen-Enzym-Immunoverfahrens.