Detection of polyketide synthetase gene expression in Karenia brevis
AssigneeUniversity of South Florida
InventorJohn H. Paul, III
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Abstract
The present invention concerns an assay for the detection of polyketide synthetase (PKS) mRNA from the red tide dinoflagellate Karenia brevis. Another aspect of the invention concerns NASBA primers and a molecular beacon that enable detection of mRNA of one of the PKS genes. In another aspect, the present invention includes an internal control RNA (IC-RNA) that enables quantitative detection of the target as well as correction for inhibition of amplification. The assay of the invention provides a rapid and specific detection of the putative toxin-producing genes of the Florida Red Tide Organism. The assay of the invention is useful for determining the virulence of red tide blooms.
Claims22
A method for detecting the presence of Karenia brevis in a sample, comprising contacting a polynucleotide region of the polyketide synthetase gene of Karenia brevis with amplification primers, wherein the amplification primers comprise one or more of SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:10;
- amplifying the polynucleotide region of the polyketide synthetase gene;
- and detecting the presence of an amplified polynucleotide, and wherein said method further comprises including an internal control (IC) nucleic acid during said amplifying step, wherein said. IC nucleic acid comprises the nucleotide sequence shown in SEQ ID NO:2, wherein the presence of the amplified polynucleotide is indicative of the presence of Karenia brevis in the sample.
A method for detecting the presence of Karenia brevis in a sample, comprising contacting a polynucleotide region of the polyketide synthetase gene of Karenta brevis with amplification primers, wherein the amplification primers comprise one or more of SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:10;
- amplifying the polynucleotide region of the polyketide synthetase gene;
- and detecting the presence of an amplified polynucleotide, and wherein said method further comprises including an internal control (IC) nucleic acid during said amplifying step, wherein said IC nucleic acid comprises the nucleotide sequence shown in SEQ ID NO:11, and wherein the sequence of “n” nucleotides does not fully or partially hybridize with the polyketide synthetase gene of Karenia brevis, wherein the presence of the amplified polynucleotide is indicative of the presence of Karenia brevis in the sample.
Dependencies are inferred from the claim text.