Development of a magnetic microplate chemifluorimmunoassay for rapid detection of bacteria and toxin in blood.
A magnetic microplate chemifluorimmunoassay (MMCIA) is described using an immunomagnetic separation and a fluorescent microplate technique for rapid detection of low-level Escherichia coli O157:H7, Bacillus subtilis var. niger spores, and Staphylococcal enterotoxin type B from whole blood. In general, the MMCIA has at least several-fold more sensitivity than the conventional enzyme-linked immunosorbent assay. In addition, the assay sensitivities using direct fluorochrome label as the reporter, or alkaline phosphatase (AP) with various assay substrates, such as pNPP and AttoPhos, were assessed.
Veridian Corporation, 5200 Springfield Pike, Suite 200, Dayton, OH 45431-1289, USA.
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D000818:Animals; D000907:Antibodies, Bacterial; D001412:Bacillus subtilis / Q000276:immunology / Q000302:isolation & purification; D001431:Bacteriological Techniques / Q000295:instrumentation / Q000379:methods* / Q000706:statistics & numerical data; D002623:Chemistry Techniques, Analytical; D004768:Enterotoxins / Q000097:blood / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000706:statistics & numerical data; D019453:Escherichia coli O157 / Q000276:immunology / Q000302:isolation & purification; D005456:Fluorescent Dyes; D015200:Fluoroimmunoassay / Q000295:instrumentation / Q000379:methods* / Q000706:statistics & numerical data; D006041:Goats; D006801:Humans; D018189:Immunomagnetic Separation; D011817:Rabbits; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012756:Sheep; D013171:Spores, Bacterial / Q000276:immunology / Q000302:isolation & purification
D000907:Antibodies, Bacterial; D004768:Enterotoxins; D005456:Fluorescent Dyes; C031818:enterotoxin B, staphylococcal