System and methods for the detection of multiple chemical compounds

• US 10,415,102 B2
• Filed: 06/05/2017
• Issued: 09/17/2019
• Est. Priority Date: 09/20/2013
• Status: Active Grant

• Current Assignee: Fredsense Technologies Corp.
• Original Assignee: Fredsense Technologies Corp.
• Inventors: Mayall, Robert Matthew, Hicks, Emily Candice, Renaud-Young, Margaret Mary-Flora, Lloyd, David Christopher, Oberding, Lisa Kara, George, Iain Fraser Scotney
• Primary Examiner(s): Noakes, Suzanne M
• Assistant Examiner(s): Lee, Jae W
• Application Number:
  US15/613,692
• Publication Number:
  US 20170335412A1
• Time in Patent Office: 834 Days
• Field of Search: None
• CPC Class Codes: C12Q 1/34 involving hydrolase
• CPC Class Codes: C12Q 1/6897 involving reporter genes operably linked to promoters
• CPC Class Codes: G01N 2333/938 acting on beta-galactose-glycoside bonds, e.g. beta-galactosidase
• CPC Class Codes: G01N 2333/942 acting on beta-1, 4-glucosidic bonds, e.g. cellulase
• CPC Class Codes: G01N 2458/30 Electrochemically active labels

Abstract

Methods that may be used for the electrochemical detection of multiple parameters, including chemical compounds. Further provided are cells that may be used in the electrochemical detection of multiple parameters, including chemical compounds, as well as a kit for the electrochemical detection of multiple parameters, including chemical compounds.

Claims

1. A diagnostic kit for simultaneously detecting two parameters comprising:

(i) a cell comprising a chimeric nucleic acid sequence comprising in the 5′

to 3′

direction of translation as operably linked components a first nucleic acid construct comprising;

(a) a first promoter operable in the cell; and

(b) a nucleic acid sequence encoding a first reporter polypeptide comprising a first hydrolase capable of hydrolyzing a first substrate comprising a first electoactive analyte and a first carrier and releasing the first electroactive analyte from the carrier; and

a second nucleic acid construct comprising;

(c) a second promoter operable in the cell; and

(d) a nucleic acid sequence encoding a second reporter polypeptide comprising a second hydrolase capable of hydrolyzing the second substrate comprising a second electroactive analytes and a second carrier and releasing the second electroactive analyte from the carrier;

(ii) a first and second substrate; and

(iii) instructions for use or storage of the kit, wherein the first and second hydrolase are independently selected from the group of hydrolases consisting of β

-D-glucosidase, β

-D galactosidase, and β

-D-glucuronidase, and wherein the first and second substrate are independently selected from the group of substrates consisting of chlorophenol red-β

-D-galactopyranoside (CPRG), para-nitrophenol-β

-D-glucuronide (PNPG) and para-diphenol-β

-D-glucopyranoside (PDPG).

2. The kit according to claim 1 wherein the first and second hydrolase are independently selected from SEQ.ID. NO:

2, SEQ.ID. NO;

4 and SEQ.ID. NO;

6.

3. The kit according to claim 1, wherein the kit is operable in conjunction with an electrical cell.

4. The kit according to claim 3 wherein the electrical cell is capable of detecting by voltammetric sweep an electrical signal facilitated by the first and second electroactive analyte in an assay mixture containing the cell and the first and second substrate.

5. The kit according to claim 1 wherein the first and second parameters are a first and second chemical compound.

6. The kit according to claim 5 wherein the first and second chemical compounds are toxic compounds present in surface water or sediments.

7. The kit according to claim 5 wherein the first and second chemical compounds are nutritional compounds used by a microorganism culture.

8. A cell comprising a chimeric nucleic acid sequence comprising in the 5′

to 3′

direction of translation as operably linked components a first nucleic acid construct comprising;

(a) a first promoter operable in the cell; and

(b) a nucleic acid sequence encoding a first reporter polypeptide comprising a first hydrolase capable of hydrolyzing a first substrate comprising a first electroactive analyte and a first carrier and releasing the first electroactive analyte from the carrier; and

a second nucleic acid construct comprising;

(c) a second promoter operable in the cell; and

(d) a nucleic acid sequence encoding a second reporter polypeptide comprising a second hydrolase capable of hydrolyzing the second substrate comprising a second electroactive analyte and a second carrier and releasing the second electroactive analyte from the carrier, wherein the first and second hydrolase are independently selected from the group of hydrolases consisting of β

-D-glucosidase, β

-D galactosidase, and β

-D-glucuronidase, and wherein the first and second substrate are independently selected from the group of substrates consisting of chlorophenol red-β

-D-galactopyranoside (CPRG), para-nitrophenol-β

-D-glucuronide (PNPG), para-aminophenol-β

-D-galactopyranoside (PAPG) and para-diphenol-β

-D-glucopyranoside (PDPG).

9. The cell according to claim 8 wherein the first and second promoter are an inducible promoter.

10. The cell according to claim 8 wherein the first and second hydrolase are independently selected from SEQ.ID. NO:

2, SEQ.ID. NO;

4 and SEQ.ID. NO;

6.