Patent ID: 8790899

Claim:
A multiplexing reaction method for detecting the presence or absence of at least one of Clostridium botulinum toxin gene A, B, E, and F in a biological sample, the method comprising: a. preparing a nucleic acid mixture comprising DNA from the biological sample; b. providing a purified polynucleic acid primer having at least 90% sequence homology to SEQ ID NO:2 and at least one isolated and purified polynucleic acid primer pair configured for use in at least one of a PCR or other amplification reaction and selected from the group consisting of sequences having at least 90% sequence homology to SEQ ID NOS: 1, 3-4, 5-6, and 7-8; c. providing at least one polynucleic acid probe selected from the group consisting of sequences having at least 90% sequence homology SEQ ID NOS 9, 10, 11, and 12, wherein each probe is labeled with a different detectable label; d. conducting an amplification reaction with a DNA polymerase using the mixture; and e. detecting hybridization of any of the at least one probe to the amplification product of step d. wherein detection is one of real time detection and delayed detection and wherein hybridization indicates the presence of at least one of toxin genes A, B, E, or F in the biological sample.