# US 8,541,006 B2 · Methods and devices for the detection of biofilm

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- Number: US 8,541,006 B2
- Document ID: US8541006B2
- Status: Granted
- Kind: B2
- Application number: 12/671,398
- Filed: 2008-07-30
- Published: 2010-11-11
- Granted: 2013-09-24
- Earliest claimed: 2007-07-30
- Assignees: University of Maryland, Baltimore; The Arizona Board of Regents, a body corporate acting for and on behalf of Northern Arizona University
- Applicants: Jeffrey G. Leid; Timothy L. Vail; Jennifer M. Kofonow; Mark E. Shirtliff; Rebecca A. Brady
- Inventors: Jeffrey G. Leid; Timothy L. Vail; Jennifer M. Kofonow; Mark E. Shirtliff; Rebecca A. Brady
- Web page: https://patentgrep.com/patent/US8541006B2

## CPC

- G01N 33/6893: {related to diseases not provided for elsewhere}
- G01N 33/54366: {Apparatus specially adapted for solid-phase testing}
- G01N 33/56938: {Staphylococcus}
- G01N 33/6854: {Immunoglobulins}
- G01N 2800/10: Musculoskeletal or connective tissue disorders
- G01N 2800/28: Neurological disorders
- 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

The present invention provides methods and kits for biofilm detection.

## Claims

### Claim 1

A method for detecting a probability for the presence of a Staphylococcus biofilm comprising:
a) contacting a test sample with one or more detectably labeled proteins,
wherein the one or more detectably labeled proteins are capable of binding antibodies present in the test sample, wherein the binding produces labeled antibodies;
b) contacting the labeled antibodies to a substrate comprising one or more immobilized biofilm markers selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:13, SEQ ID NO: 43 and a polypeptide comprising one or more antigenic fragments thereof; and
c) detecting binding of the labeled antibodies to the one or more immobilized biofilm markers, wherein binding indicates a probability for the presence of the Staphylococcus biofilm in the test sample.

### Claim 2 (depends on 1)

The method of claim 1 wherein the contacting the labeled antibodies to a substrate comprises allowing the labeled antibodies to migrate along the substrate prior to contacting the one or more immobilized biofilm markers.

### Claim 3 (depends on 1)

The method of claim 1 wherein the biofllm comprises Staphylococcus aureus.

### Claim 4 (depends on 3)

The method of claim 3 wherein the Staphylococcus aureus comprises methicillin-resistant Staphylococcus aureus.

### Claim 5

A method for diagnosing a probability for the presence of a Staphylococcus biofilm related disease in a subject, comprising:
a) contacting a test sample from the subject with one or more detectably labeled proteins, wherein the one or more detectably labeled proteins are capable of binding antibodies present in the test sample, wherein the binding produces labeled antibodies;
b) contacting the labeled antibodies to a substrate comprising one or more immobilized biofilm markers selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:13, SEQ ID NO: 43 and a polypeptide comprising one or more antigenic fragments thereof; and
c) detecting binding of the labeled antibodies to the one or more immobilized biofilm markers, wherein binding indicates a probability for the presence of a Staphylococcus biofilm related disease in the subject.

### Claim 6 (depends on 5)

The method of claim 5, wherein the biofilm related disease is osteomyelitis.

### Claim 7

Biofilm detection substrates for detecting a probability for the presence of a Staphylococcus biofilm comprising:
a) a test well comprising one or more detectably labeled proteins, wherein the one or more detectably labeled proteins are capable of binding to biofilm antibodies present in a test sample; and
b) one or more immobilized Staphylococcus biofilm markers capable of binding to labeled antibodies selected from the group consisting of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:13, SEQ ID NO: 43 and a polypeptide comprising one or more antigenic fragments thereof.

### Claim 8 (depends on 7)

A kit for detecting a probability for the presence of a Staphylococcus biofilm comprising the substrates of claim 7, wherein the one or more detectably labeled proteins of a) are capable of binding to biofilm antibodies present in a test sample, wherein the test sample is a bodily fluid sample from a patient.

### Claim 9 (depends on 1)

The method of claim 1, wherein the one or more immobilized biofilm markers of b) is SEQ ID NO:1 or a polypeptide comprising one or more antigenic fragments thereof.

### Claim 10 (depends on 1)

The method of claim 1, wherein the one or more immobilized biofilm markers of b) is SEQ ID NO:2 or a polypeptide comprising one or more antigenic fragments thereof.

### Claim 11 (depends on 1)

The method of claim 1, wherein the one or more immobilized biofilm markers of b) is SEQ ID NO:3 or a polypeptide comprising one or more antigenic fragments thereof.

### Claim 12 (depends on 1)

The method of claim 1, wherein the one or more immobilized biofilm markers of b) is SEQ ID NO:13 or a polypeptide comprising one or more antigenic fragments thereof.

### Claim 13 (depends on 1)

The method of claim 1, wherein the one or more immobilized biofilm markers of b) is SEQ ID NO:43 or a polypeptide comprising one or more antigenic fragments thereof.

### Claim 14 (depends on 5)

The method of claim 5, wherein the one or more immobilized biofilm markers of b) is SEQ ID NO:1 or a polypeptide comprising one or more antigenic fragments thereof.

### Claim 15 (depends on 5)

The method of claim 5, wherein the one or more immobilized biofilm markers of b) is SEQ ID NO:2 or a polypeptide comprising one or more antigenic fragments thereof.

### Claim 16 (depends on 5)

The method of claim 5, wherein the one or more immobilized biofilm markers of b) is SEQ ID NO:3 or a polypeptide comprising one or more antigenic fragments thereof.

### Claim 17 (depends on 5)

The method of claim 5, wherein the one or more immobilized biofilm markers of b) is SEQ ID NO:13 or a polypeptide comprising one or more antigenic fragments thereof.

### Claim 18 (depends on 5)

The method of claim 5, wherein the one or more immobilized biofilm markers of b) is SEQ ID NO:43 or a polypeptide comprising one or more antigenic fragments thereof.

### Claim 19 (depends on 7)

The biofilm detection substrates of claim 7, wherein the one or more immobilized biofilm markers of b) is SEQ ID NO:1 or a polypeptide comprising one or more antigenic fragments thereof.

### Claim 20 (depends on 7)

The biofilm detection substrates of claim 7, wherein the one or more immobilized biofilm markers of b) is SEQ ID NO:2 or a polypeptide comprising one or more antigenic fragments thereof.