Method for detecting ammonia-oxidizing bacteria
AssigneeAquaria, Inc.
InventorTimothy A. Hovanec
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Abstract
Described herein are ammonia-oxidizing bacteria. Particular bacteria of the present invention are tolerant of freshwater environments, saltwater environments or both. Furthermore, in various embodiments, various bacteria of the present invention are capable of surviving a freeze-drying process, and may remain viable thereafter. Methods for detecting the bacteria of the present invention are also provided. Such methods may be effected by any conventional methology, such as with a DNA chip.
Claims32
A method for detecting and determining the quantity of bacteria that oxidize ammonia to nitrite in a medium, wherein the 16S rDNA of the bacteria includes a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3 and SEQ ID NO:4, comprising:
- providing a detectably labeled probe including a nucleotide sequence selected from the group consisting of SEQ ID NO:5 and SEQ ID NO:8;
- isolating total DNA from the medium;
- exposing the isolated total DNA to the detectably labeled probe under conditions under which the probe hybridizes to only the nucleic acid of bacteria having 16S rDNA including the nucleotide sequence;
- and
- detecting and measuring the amount of hybridized probe,
- wherein the presence of hybridized probe is indicative of the presence of bacteria that oxidize ammonia to nitrite and the amount of hybridized probe is indicative of the quantity of said bacteria that oxidize ammonia to nitrite in said medium.
A method for detecting and determining the quantity of bacteria that oxidize ammonia to nitrite in a medium, wherein the 16S rDNA of the bacteria includes a nucleotide sequence selected from the group consisting of SEQ ID NO:18, SEQ ID NO:19 and SEQ ID NO:20 comprising:
- providing a detectably labeled probe including a nucleotide sequence as set forth in SEQ ID NO:21;
- isolating total DNA from the medium;
- exposing the isolated total DNA to the detectably labeled probe under conditions under which the probe hybridizes to only the nucleic acid of bacteria having 16S rDNA including the nucleotide sequence;
- and
- detecting and measuring the amount of hybridized probe,
- wherein the presence of hybridized probe is indicative of the presence of bacteria that oxidize ammonia to nitrite and the amount of hybridized probe is indicative of the quantity of said bacteria that oxidize ammonia to nitrite in said medium.
A method for detecting and determining the quantity of bacteria that oxidize ammonia to nitrite in a medium, wherein the 16S rDNA of the bacteria includes a nucleotide sequence selected from the group consisting of:
- a nucleotide sequence that has greater than 98% identity over the full length thereof to SEQ ID NO:3, a nucleotide sequence that has greater than 98% identity over the full length thereof to SEQ ID NO:4, a nucleotide sequence that has at least 96% identity over the full length thereof to SEQ ID NO:1 and a nucleotide sequence that has at least 96% identity over the full length thereof to SEQ ID NO:2, comprising:
- providing a detectably labeled probe that has at least 96% identity over the full length thereof to a nucleotide sequence selected from the group consisting of SEQ ID NO:5 and SEQ ID NO:8;
- isolating total DNA from the medium;
- exposing the isolated total DNA to the detectably labeled probe under conditions under which the probe hybridizes to only the nucleic acid of bacteria having 16S rDNA including the nucleotide sequence;
- and
- detecting and measuring the amount of hybridized probe,
- wherein the presence of hybridized probe is indicative of the presence of bacteria that oxidize ammonia to nitrite and the amount of hybridized probe is indicative of the quantity of said bacteria that oxidize ammonia to nitrite in said medium.
A method for detecting and determining the quantity of bacteria that oxidize ammonia to nitrite in a medium, wherein the 16S rDNA of the bacteria includes a nucleotide sequence selected from the group consisting of:
- a nucleotide sequence that has at least 96% identity over the full length thereof to SEQ ID NO:18, at least 96% identity over the full length thereof to SEQ ID NO:19 and at least 96% identity over the full length thereof to SEQ ID NO:20, comprising:
- providing a detectably labeled probe that has at least 96% identity over the full length thereof to a nucleotide sequence as set forth in SEQ ID NO:21;
- isolating total DNA from the medium;
- exposing the isolated total DNA to the detectably labeled probe under conditions under which the probe hybridizes to only the nucleic acid of bacteria having 16S rDNA including the nucleotide sequence;
- and
- detecting and measuring the amount of hybridized probe,
- wherein the presence of hybridized probe is indicative of the presence of bacteria that oxidize ammonia to nitrite and the amount of hybridized probe is indicative of the quantity of said bacteria that oxidize ammonia to nitrite in said medium.
A DNA chip comprising:
- a solid substrate;
- and
- a probe including a nucleotide sequence selected from the group consisting of SEQ ID NO:5 and SEQ ID NO:8 configured upon the solid substrate.
A DNA chip comprising:
- a solid substrate;
- and
- a probe including a nucleotide sequence as set forth in SEQ ID NO:21 configured upon the solid substrate.
Dependencies are inferred from the claim text.