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US 2002/0058294 A1Published application
Method for detecting clostridium botulinum neurotoxin serotypes A, B, E and F in a sample
OwnerMark A. Poli, Victor R. Riversinventors
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Abstract
Sensitive and specific enzyme-linked immunosorbent assays which detect Clostridium botulinum neurotoxins serotypes A, B, E, and F in a sample are described. The assay is based upon affinity-purified antibodies directed against the C-fragments of each toxin. These assays demonstrate sensitivity close to that of the mouse bioassay without the use of animals and in a much simpler format than other assays of similar sensitivity.
Claims10
A method for detecting BoNT antigen in a sample comprising:
- (a) reacting a sample suspected of having an antigen with a first antibody, Ab(A);
- (b) washing the reaction mixture to remove unreacted antibody;
- (c) reacting the Ab(A) linked antigen with a second antibody which is conjugated with biotin, Ab(B);
- (d) washing the reaction mixture to remove unreacted antibody;
- (e) reacting Ab(A)-antigen-Ab(B) with avidin linked to a chromogenic enzyme;
- (f) washing the reaction mixture;
- (g) reacting the chromogenic enzyme;
- (h) measuring optical density of reaction and calculating concentration of antigen using a standard curve.
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A test kit for the assay of BoNT antigen comprising:
- one or more solutions containing a known concentration of free BoNT or complexed BoNT to serve as a standard;
- a solution of a anti-BoNT antibody, and biotinylated anti-BoNT antibody affinity purified to the C-fragment of BoNT;
- a chromogen which changes color or shade by action with biotin;
- and, optionally,
- a solid support.
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Dependencies are inferred from the claim text.