# US 10,145,833 B2 · Detection method

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- Number: US 10,145,833 B2
- Document ID: US10145833B2
- Status: Granted
- Kind: B2
- Application number: 10/468,691
- Filed: 2002-02-21
- Published: 2004-06-17
- Granted: 2018-12-04
- Earliest claimed: 2001-02-23
- Assignees: Conidia Bioscience Limited
- Applicants: Joan Kelley
- Inventors: Joan Kelley
- Web page: https://patentgrep.com/patent/US10145833B2

## CPC

- G01N 33/22: Fuels; Explosives
- G01N 33/56961: {Plant cells or fungi}
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

A method of analysis of a hydrocarbon fuel for the presence of a micro-organism comprises contacting a fuel sample with an aqueous diluent and with an antibody reactive with the micro-organism, or reactive with a metabolite or breakdown product produced by the micro-organism, to detect the presence or absence of the micro-organism.

## Claims

### Claim 1

A method of analysis of a sample of a liquid hydrocarbon fuel for the presence of Hormoconis resinae, the method comprising the steps of:
1) mixing the liquid fuel sample with an aqueous diluent containing an emulsifier;
2) contacting the mixture or one phase thereof with an antiserum comprising antibodies raised against a homogenized preparation of Hormoconis resinae, wherein the Hormoconis resinae in the homogenized preparation was grown in contact with the liquid hydrocarbon fuel; and
3) detecting the presence or absence of bound antibodies in the sample,
wherein the method provides a result within two hours from step 1, and wherein the detection of bound antibodies indicates the presence of Hormoconis resinae which is actively growing in liquid hydrocarbon fuel.

### Claim 2 (depends on 1)

The method of claim 1, wherein the method comprises vigorously mixing the sample of fuel and the aqueous diluent before contacting the mixture with the antibodies.

### Claim 3 (depends on 1)

The method of claim 1, wherein the emulsifier is a non-ionic detergent.

### Claim 4 (depends on 3)

The method of claim 3, wherein the emulsifier is polyoxyethylene (20) sorbitan monolaurate.

### Claim 5 (depends on 4)

The method of claim 4, wherein the emulsifier is used at a concentration of from 1 to 10% by volume.

### Claim 6 (depends on 1)

The method of claim 1, wherein the hydrocarbon fuel is light-middle distillate fuel having a boiling point below 390° C.

### Claim 7 (depends on 1)

The method of claim 1, wherein the fuel sample is not filtered prior to the step of contacting the mixture or one phase thereof with the antibodies.

### Claim 8 (depends on 1)

The method of claim 1, wherein the antibodies are not incubated with a filter used to filter the sample of the liquid hydrocarbon fuel.

### Claim 9

A kit for analyzing a sample of a liquid hydrocarbon fuel for the presence of Hormoconis resinae, the kit comprising:
1) a homogenized preparation of Hormoconis resinae, wherein the Hormoconis resinae in the homogenized preparation was grown in contact with liquid hydrocarbon fuel;
2) an antiserum comprising antibodies raised against a homogenized preparation of Hormoconis resinae, wherein the Hormoconis resinae in the homogenized preparation was grown in contact with the liquid hydrocarbon fuel; and
3) an aqueous diluent containing an emulsifier.

### Claim 10 (depends on 9)

The kit of claim 9, further including a substrate to which the antibodies are immobilized.

### Claim 11 (depends on 10)

The kit of claim 10, wherein the substrate is a microtiter plate.

### Claim 12 (depends on 9)

The kit of claim 9, further including a lateral flow device to which the antibodies are immobilized.