Compositions, products, methods and systems to monitor water and other ecosystems

• US 8,383,342 B2
• Filed: 10/07/2011
• Issued: 02/26/2013
• Est. Priority Date: 04/24/2002
• Status: Expired due to Fees

• Current Assignee: University of North Carolina At Greensboro (University of North Carolina System)
• Original Assignee: University of North Carolina At Greensboro (University of North Carolina System)
• Inventors: Rublee, Parke A., Henrich, Vincent C. III, Marshall, Michael McClain
• Primary Examiner(s): CROW, ROBERT THOMAS
• Application Number:
  US13/269,041
• Publication Number:
  US 20120021429A1
• Time in Patent Office: 508 Days
• Field of Search: 435/6, 435/6.1, 435/283.1, 435/287.1, 435/287.2
• US Class Current: 435/6.1
• CPC Class Codes: C12Q 1/689 for bacteria
• CPC Class Codes: C12Q 2600/16 Primer sets for multiplex assays

Abstract

Disclosed are compositions, products, methods and systems for monitoring ecosystems, such as bodies of water, for a parameter of the ecosystems, such as the presence or absence of mercury. In one embodiment, the product may include a plurality of oligonucleotides immobilized at known locations on a substrate as an array, such that each location on the array is an oligonucleotide having a sequence derived from a single, predetermined operational taxonomic unit (OTU) and wherein at least one sequence on the array is associated with the presence or absence of mercury. The sequences immobilized on the array may be from known, or unknown organisms. Also disclosed are methods for identifying and isolating bioindicators diagnostic of ecosystem parameters, such as whether mercury is present. The compositions, products, methods and systems of the invention may be used for rapid, and continual monitoring of ecosystems for parameters of interest, such as the presence or absence of mercury.

Claims

1. A composition comprising an isolated oligonucleotide, wherein the sequence of the isolated oligonucleotide is specific to a single operational taxonomic unit in an aquatic ecosystem, and wherein the oligonucleotide comprises a bioindicator for a parameter in the aquatic ecosystem, wherein the parameter is mercury levels in the ecosystem, and wherein the isolated oligonucleotide comprises the sequence as set forth in SEQ ID NO:

355 or SEQ ID NO;

356, where SEQ ID NO;

356 is the reverse complement of SEQ ID NO;

355.

2. The composition of claim 1, further comprising at least one additional oligonucleotide having the sequence as set forth in SEQ ID NOS:

329-340 or SEQ ID NOS;

351-354 or SEQ ID NOS;

357-370.

3. The composition of claim 1, further comprising two oligonucleotides derived from a single genomic target sequence such that the two oligonucleotides can be used as primers in a polymerase chain reaction to amplify DNA from the genomic target sequence.

4. The composition of claim 3, comprising at least 6 distinct oligonucleotides, the 6 distinct oligonucleotides each comprising at least one of the sequences as set forth in SEQ ID NOS:

341-350 or SEQ ID NOS;

371-388, wherein the at least 6 distinct oligonucleotides can be used as primers in a polymerase chain reaction to amplify DNA from a plurality of genomic target sequences.

5. The composition of claim 3, comprising at least 10 distinct oligonucleotides, the 10 distinct oligonucleotides each comprising at least one of the sequences as set forth in SEQ ID NOS:

341-350 or SEQ ID NOS;

371-388, wherein the at least 10 distinct oligonucleotides can be used as primers in a polymerase chain reaction to amplify DNA from a plurality of genomic target sequences.

6. The composition of claim 3, wherein the two oligonucleotides have the sequences as set forth in SEQ ID NOS:

375 and 376.

7. The composition of claim 1, further comprising at least three additional oligonucleotides having the sequence as set forth in SEQ ID NOS:

329-340 or SEQ ID NOS;

351-354 or SEQ ID NOS;

357-370.

8. The composition of claim 1, further comprising at least five additional oligonucleotides having the sequence as set forth in SEQ ID NOS:

329-340 or SEQ ID NOS;

351-354 or SEQ ID NOS;

357-370.

9. A product for monitoring an aquatic ecosystem, the product comprising a plurality of individual locations each comprising an isolated oligonucleotide that is a bioindicator for a parameter in the aquatic ecosystem, the oligonucleotide having a nucleic acid sequence that is derived from a single operational taxonomic unit in an aquatic ecosystem, and wherein at least one of the oligonucleotides has the sequence as set forth in SEQ ID NO:

355 or SEQ ID NO;

356, where SEQ ID NO;

356 is the reverse complement of SEQ ID NO;

355.

10. The product of claim 9, further comprising two oligonucleotides derived from a single genomic target sequence such that the two oligonucleotides can be used as primers in a polymerase chain reaction to amplify DNA from the genomic target sequence.

11. The product of claim 10, wherein the two oligonucleotides each comprise at least one of the sequences as set forth in SEQ ID NOS:

341-350 or SEQ ID NOS;

371-388.

12. The product of claim 11, wherein the two oligonucleotides have the sequences as set forth in SEQ ID NO:

375 and 376.

13. The product of claim 10, further comprising at least six additional oligonucleotides each comprising one of the sequences as set forth in SEQ ID NOS:

341-350 or SEQ ID NOS;

371-388, wherein the six additional oligonucleotides can be used as primers in a polymerase chain reaction to amplify DNA from the genomic target sequence.

14. The product of claim 10, further comprising at least ten additional oligonucleotides comprising one of the sequences as set forth in SEQ ID NOS:

341-350 or SEQ ID NOS;

371-388, wherein the ten additional oligonucleotides can be used as primers in a polymerase chain reaction to amplify DNA from the genomic target sequence.

15. The product of claim 9, further comprising at least one additional location comprising an oligonucleotide having the sequence as set forth in SEQ ID NOS:

329-340 or SEQ ID NOS;

351-354 or SEQ ID NOS;

357-370.

16. The product of claim 9, further comprising at least three additional locations each comprising an oligonucleotide having the sequence as set forth in SEQ ID NOS:

329-340 or SEQ ID NOS;

351-354 or SEQ ID NOS;

357-370.

17. The product of claim 9, wherein the oligonucleotides are immobilized at each of the locations.

18. The product of claim 9, wherein the parameter is the level of mercury in the ecosystem.

19. The product of claim 9, further comprising at least five additional locations each comprising an oligonucleotide having the sequence as set forth in SEQ ID NOS:

329-340 or SEQ ID NOS;

351-354 or SEQ ID NOS;

357-370.