# US 8,541,188 B2 · Differential immunoassay for PRRS vaccine antibody

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- Number: US 8,541,188 B2
- Document ID: US8541188B2
- Status: Granted
- Kind: B2
- Application number: 13/298,776
- Filed: 2011-11-17
- Published: 2012-03-08
- Granted: 2013-09-24
- Earliest claimed: 2005-12-02
- Assignees: Iowa State University Research Foundation, Inc.
- Applicants: Kyoung-Jin Yoon; Wai-Hong Wu
- Inventors: Kyoung-Jin Yoon; Wai-Hong Wu
- Web page: https://patentgrep.com/patent/US8541188B2

## CPC

- G01N 33/56983: {Viruses}
- G01N 2333/08: RNA viruses
- 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 relates to immunoassays for serologically differentiating animals naturally infected with PRRS virus from animals vaccinated against PRRS. The immunoassays provide detection of at least a portion of the N terminal region of the 2b portion of PRRSV. The immunoassay is preferably an enzyme-linked immunosorbent assay (ELISA).

## Claims

### Claim 1

A method of determining whether a pig has received a commercial PRRSV modified live attenuated vaccine (MLV) comprising:
a. obtaining a biological sample from an individual;
b. contacting the biological sample with an antibody that binds to the N terminal region of the 2b protein of PRRSV to form a complex when there is an aspartic acid at position 9; wherein the position number of said PRRSV corresponds to the position number in SEQ ID NO:40; and
c. assaying for the presence of said complex, wherein an aspartic acid at position 9 indicates that said pig has not received a commercial PRRSV modified live attenuated vaccine (MLV) and a tyrosine or glycine at said position indicates that said pig has received said MLV.

### Claim 2 (depends on 1)

The method of claim 1 wherein the PRRSV is free or bound to a carrier molecule or solid support.

### Claim 3 (depends on 1)

The method of claim 1 wherein the antibody is selected from the group consisting of a polyclonal antibody and a monoclonal antibody.

### Claim 4 (depends on 1)

The method of claim 1 wherein the biological sample is selected from the group consisting of urine, whole blood, plasma, serum, saliva, semen, stool, sputum, cerebral spinal fluid, tears, mucus, tissue, and tissue extract.

### Claim 5 (depends on 1)

The method of claim 1 wherein the antibody is linked to or incorporates a label.

### Claim 6 (depends on 1)

The method of claim 1 wherein the assaying step uses an immunoassay selected from the group consisting of EMIT, radio immunoassay, fluorescent immunoassay, enzyme channeling techniques, and ELISA.

### Claim 7 (depends on 1)

The method of claim 1 wherein the assaying step uses ELISA-based immunoenzymatic detection.

### Claim 8

A method of determining whether a pig has received a commercial PRRSV modified live attenuated vaccine (MLV) in a biological sample comprising:
a. contacting the biological sample with an antibody or fragment thereof that binds to the N terminal region of the 2b protein of PRRSV to form a complex when there is an aspartic acid at position 9 corresponding to position 9 of SEQ ID NO:40;
b. assaying for the presence of said complex; and
c. determining the amino acid present at position 9 of the 2b protein of PRRSV in said sample, wherein an aspartic acid at position 9 indicates that a pig has not received a commercial PRRSV Modified live attenuated vaccine (MLV) and a tyrosine or glycine at said position indicates that said pig has received said commercial PRRSV modified live attenuated vaccine (MLV).

### Claim 9 (depends on 8)

The method of claim 8 wherein the antibody or fragment thereof is a polyclonal antibody.