Abstract
We describe an ultrasensitive immunoassay for detecting biotoxins that uses a liposome with encapsulated DNA reporters, and ganglioside receptors embedded in the bilayer, as the detection reagent. After immobilization of the target biotoxin by a capture antibody and co-binding of the detection reagent, the liposomes are ruptured to release the reporters, which are quantified by real-time polymerase chain reaction. The new assays for cholera and botulinum toxins are several orders of magnitude more sensitive than current detection methods. A single 96-well microtiter plate can analyze ∼20 specimens, including calibration standards and controls, with all measurements conducted in triplicate. Using pre-coated and blocked microtiter plates, and pre-prepared liposome reagents, a liposome polymerase chain reaction assay can be carried out in about 6 h.
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This work was supported by Army Medical Research and Material Command grant DAMD17-02-1-0178.
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Mason, J., Xu, L., Sheng, Zm. et al. Liposome polymerase chain reaction assay for the sub-attomolar detection of cholera toxin and botulinum neurotoxin type A. Nat Protoc 1, 2003–2011 (2006). https://doi.org/10.1038/nprot.2006.331
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DOI: https://doi.org/10.1038/nprot.2006.331
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