# US 2013/0130274 A1 · Detection method

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- Number: US 2013/0130274 A1
- Document ID: US20130130274A1
- Status: Published application
- Kind: A1
- Application number: 13/736,541
- Filed: 2013-01-08
- Published: 2013-05-23
- Earliest claimed: 2001-02-23
- Assignees: Conidia Bioscience Limited
- Inventors: Joan Kelly
- Web page: https://patentgrep.com/patent/US20130130274A1

## 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 of 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) emulsifying the liquid fuel sample by vigorously mixing the liquid fuel sample with an aqueous diluent containing an emulsifier;
2) contacting the emulsified mixture with antibodies specific for an antigen that is a metabolite produced by Hormoconis resinae which is recoverable from liquid hydrocarbon fuel when Hormoconis resinae is 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 according to claim 1, wherein the emulsifier is a non-ionic detergent.

### Claim 3 (depends on 3)

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

### Claim 4 (depends on 3)

The method according to claim 3, wherein the emulsifier is used at a concentration of 1-10% by volume.

### Claim 5 (depends on 1)

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

### Claim 6

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) antibodies specific for an antigen recoverable from the liquid hydrocarbon fuel after the Hormoconis resinae was grown in contact with the liquid hydrocarbon fuel; and
3) an aqueous diluent containing an emulsifier.

### Claim 7 (depends on 6)

The kit of claim 6, further including a substrate to which the antibodies are immobilized.

### Claim 8 (depends on 7)

The kit of claim 7, wherein the substrate is a microtiter plate.

### Claim 9 (depends on 6)

The kit of claim 6, further including a lateral flow device to which the antibodies are immobilized.

### Claim 10 (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 11 (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.