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

• US 8,048,623 B1
• Filed: 08/04/2008
• Issued: 11/01/2011
• 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: Henrich, Vincent C. III, Rublee, Parke A., Marshall, Michael McClain
• Primary Examiner(s): CROW, ROBERT THOMAS
• Application Number:
  US12/221,541
• Time in Patent Office: 1,184 Days
• Field of Search: 435/6, 435/91.2, 435/287.2, 536/23.1, 536/24.1
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/6888 for detection or identification of organisms
• CPC Class Codes: C12Q 2565/501 being an array of oligonucleotides

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 method to monitor a parameter of an aquatic ecosystem comprising the steps of:

(a) obtaining a sample from an aquatic ecosystem;

(b) using at least one oligonucleotide specific to a single operational taxonomic unit to determine whether a nucleic acid sequence specific to the operational taxonomic unit is present in the sample wherein the at least one oligonucleotide specific to the single operational taxonomic unit comprises SEQ ID NO;

355 or SEQ ID NO;

356; and

(c) correlating detection of the nucleic acid sequence specific to the operational taxonomic unit to a parameter of the ecosystem wherein the parameter is mercury.

2. The method of claim 1, further comprising generating a plurality of distinct oligonucleotides each specific to a single operational taxonomic unit in at least one aquatic ecosystem and determining whether nucleic acid sequences specific to each of the operational taxonomic units are present in the sample.

3. The method of claim 1, further comprising identifying the at least one oligonucleotide specific to a single operational taxonomic unit in a first aquatic ecosystem and using the at least one oligonucleotide specific to a single operational taxonomic unit to determine whether nucleic acid sequences specific to the operational taxonomic unit are present in a second aquatic ecosystem.

4. The method of claim 1, wherein step (b) comprises using two oligonucleotides as primers in a polymerase chain reaction to amplify DNA from a single genomic target sequence wherein the amplified product comprises the at least one oligonucleotide specific to a single operational taxonomic unit.

5. The method of claim 2, wherein step (b) comprises using a plurality of paired oligonucleotides as primers in a plurality of separate polymerase chain reactions to amplify DNA from a plurality of genomic target sequences wherein the amplified product comprises at least some of the plurality of the distinct oligonucleotides each specific to a single operational taxonomic unit.

6. The method of claim 1, wherein the at least one oligonucleotide specific to the single operational taxonomic unit further comprises an oligonucleotide comprising SEQ ID NO:

357 or 358.

7. The method of claim 2, wherein the plurality of distinct oligonucleotides comprises an array of individual locations, each location comprising at least one oligonucleotide specific to a single operational taxonomic unit.

8. The method of claim 1, wherein the at least one oligonucleotide specific to the single operational taxonomic unit further comprises an oligonucleotide having the sequence as set forth in any of SEQ ID NOs:

329-340 or SEQ ID NOs;

351-354 or SEQ ID NOs;

359-370.

9. The method of claim 1, wherein step (b) comprises using two oligonucleotides as primers in a polymerase chain reaction to amplify DNA from a single genomic target sequence wherein the amplified product comprises the oligonucleotide specific to a single operational taxonomic unit and wherein the primers comprise oligonucleotides comprising a first primer having the sequence as set forth in SEQ ID NO:

375 and a second primer having the sequence as set forth in SEQ ID NO;

376.

10. The method of claim 8, wherein step (b) comprises using two oligonucleotides as primers in a polymerase chain reaction to amplify DNA from a single genomic target sequence wherein the amplified product comprises the oligonucleotide specific to a single operational taxonomic unit and wherein the primers comprise oligonucleotides comprising the sequence as set forth in any of SEQ ID NOs:

341-350 or SEQ ID NOs;

371-374 or SEQ ID NOs;

377-388.