Detection of saxitoxin-producing dinoflagellates

Patent metadata

Field	Value
Publication number	US-10947601-B2
Application number	US-201615370752-A
Country	US
Kind code	B2
Filing date	Dec 6, 2016
Priority date	May 16, 2011
Publication date	Mar 16, 2021
Grant date	Mar 16, 2021

Abstract

The invention generally relates to the field of saxitoxins and the identification of microorganisms capable of producing them. More specifically, the invention relates to the identification of genes encoding saxitoxin in dinoflagellates, and methods for the specific detection of dinoflagellates that are producers of saxitoxins.

First claim

The invention claimed is:

1. A method comprising:
obtaining a sample for use in the method; and
detecting whether a saxitoxin-producing dinoflagellate is present in the sample by contacting the sample with a nucleic acid primer specific for a dinoflagellate saxitoxin A catalytic domain polynucleotide sequence or an antibody specific for a dinoflagellate saxitoxin A catalytic domain polypeptide, wherein the saxitoxin A catalytic domain is selected from: saxitoxin A1 catalytic domain, saxitoxin A4 catalytic domain, or a fragment of a saxitoxin A1 or A4 catalytic domain, and
wherein:
the saxitoxin A4 catalytic domain sequence consists of a nucleotide sequence having 95% or more similarity with nucleotides 3115-4121 of the polynucleotide sequence set forth in SEQ ID NO: 3, or a nucleotide sequence having 95% or more similarity with nucleotides 3597-3721 of the polynucleotide sequence set forth in SEQ ID NO: 3, and
the saxitoxin A1 catalytic domain sequence consists of a nucleotide sequence having 95% or more similarity with nucleotides 160-1821 of the polynucleotide sequence set forth in SEQ ID NO: 1, or a nucleotide sequence having 95% or more similarity with nucleotides 277-2022 of the polynucleotide sequence set forth in SEQ ID NO: 3.

2. The method according to claim 1 , wherein the polynucleotide comprises a saxitoxin A nucleotide sequence selected from a nucleotide sequence having 95% or more similarity with any one of SEQ ID NOS: 224-227, 230-242 and 247.

3. The method according to claim 1 , wherein the saxitoxin A catalytic domain is a saxitoxin A4 catalytic domain sequence or fragment thereof consisting of a nucleotide sequence having 95% or more similarity with nucleotides 3115-4121 of the polynucleotide sequence set forth in SEQ ID NO: 3, or a nucleotide sequence having 95% or more similarity with nucleotides 3597-3721 of the polynucleotide sequence set forth in SEQ ID NO: 3.

4. The method according to claim 1 , wherein the saxitoxin A catalytic domain is a saxitoxin A1 catalytic domain sequence consisting of a nucleotide sequence having 95% or more similarity with nucleotides 160-1821 of the polynucleotide sequence set forth in SEQ ID NO: 1, or a nucleotide sequence having 95% or more similarity with nucleotides 277-2022 of the polynucleotide sequence set forth in SEQ ID NO: 3.

5. The method according to claim 1 , wherein said detecting comprises amplification of polynucleotides from the sample by polymerase chain reaction and said polymerase chain reaction utilises one or more primers comprising a sequence set forth in any one of SEQ ID NOs: 198-199, 200-211, 228-229, and 243-244, or a fragment of any one of those sequences.

6. The method according to claim 1 , wherein the polypeptide comprises:
(i) a saxitoxin A1 catalytic domain amino acid sequence having 95% or more similarity with residues 1-554 of SEQ ID NO: 2 or a saxitoxin A1 catalytic domain amino acid sequence having 95% or more similarity with residues 1-582 of SEQ ID NO: 4; or
(ii) a saxitoxin A4 catalytic domain amino acid sequence having 95% or more similarity with residues 947-1281 of SEQ ID NO: 4.

7. The method according to claim 1 , wherein the saxitoxin-producing dinoflagellate is from the Alexandrium, Pyrodinium or Gymnodinium genus.

8. The method according to claim 7 , wherein the saxitoxin-producing dinoflagellate is selected from the group consisting of A. catenella, A. fundyense, A lusitanicum, A. minutum, A. ostenfeldii, A. tamarense, G. catenatum and P. bahamense var compressum.

Assignees

• Newsouth Innovations Pty Ltd
• Univ Oslo
• Ewsouth Innovations Pty Ltd

Inventors

• Neilan Brett A
• Murray Shauna Ann
• Stuken Anke
• Jakobsen Kjetill S
• Orr Russel J S
• Kellmann Ralf

Classifications

• C12Q1/6893 Primary

  for protozoa · CPC title

• C12N15/52

  Genes encoding for enzymes or proenzymes · CPC title

• C12Q1/04

  Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor {(C12Q1/6897 takes precedence)} · CPC title

• C12P17/182

  Heterocyclic compounds containing nitrogen atoms as the only ring heteroatoms in the condensed system (alloxazine or isoalloxazine, e.g. riboflavine C12P25/00) · CPC title

• C07K14/44

  from protozoa · CPC title