Methods for Diagnosing Lyme Disease

• US 20130273572A1
• Filed: 09/27/2011
• Published: 10/17/2013
• Est. Priority Date: 09/27/2010
• Status: Active Grant

• Current Assignee: Cornell University
• Original Assignee: Cornell University
• Inventors: Wagner, Bettina
• Granted Patent: US 8,946,393 B2
• Time in Patent Office: Days
• US Class Current: 435/7.32
• CPC Class Codes: C07K 14/20 from Spirochaetales (O), e.g. Treponema, Leptospira
• CPC Class Codes: G01N 2333/20 from Spirochaetales (O), e.g. Treponema, Leptospira
• CPC Class Codes: G01N 33/56911 Bacteria
• CPC Class Codes: G01N 33/6854 Immunoglobulins
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

A method for diagnosing Lyme disease status in a mammal is provided. The method entails, in a biological sample obtained or derived from a mammal, determining antibodies to Borrelia burgdorferi (B. burgdorferi) outer surface proteins (Osp) OspA, OspC, and OspF. Based upon determining the OspA, OspC, and OspF antibodies, the mammal can be diagnosed as vaccinated, not vaccinated, infected or not infected with B. burgdorferi. Mammals that have early, intermediate or chronic B. burgdorferi infection can also be identified. The method is particularly suited for use with horses and dogs. Isolated or recombinant B. burgdorferi antigens and compositions that contain them are also provided.

Claims

1. A method for diagnosing Lyme disease status in a mammal comprising, in a biological sample obtained or derived from the mammal, determining presence or absence of antibodies to Borrelia burgdorferi (B. burgdorferi) outer surface proteins (Osp) OspA, OspC, and OspF, and based on the presence or absence of the antibodies, identifying the mammal as infected or not infected with B. burgdorferi.

2. The method of claim 1, wherein the mammal is a horse or a dog.

3. The method of claim 1, wherein the only antibodies to B. burgdorferi antigens determined in the method are antibodies to B. burgdorferi Osp A, Osp C, and Osp F.

4. The method of claim 3, wherein the mammal is identified as not infected with B. burgdorferi by determining an absence of the antibodies to the OspA, OspC and OspF antigens.

5. The method of claim 1, further comprising determining that the animal has an early, intermediate or chronic B. burgdorferi infection based on the determining the presence or the absence of the antibodies.

6. The method of claim 1, further comprising determining whether the mammal has been vaccinated against B. burgdorferi infection based on the determining the presence or the absence of the antibodies.

7. The method of claim 1, wherein the determining the presence or the absence of the antibodies is determined using a lateral flow device.

8. The method of claim 1, wherein the determining the presence or the absence of the antibodies is determined using a fluorescent bead-based multiplex assay.

9. A method for diagnosing Lyme disease status in a mammal comprising, in a biological sample obtained or derived from the mammal, determining a test level of antibodies to Borrelia burgdorferi (B. burgdorferi) outer surface proteins (Osp) Osp A, Osp C, and Osp F, and based upon a comparison of the test level of the Osp A, Osp C, and Osp F antibodies to a reference, identifying the mammal as vaccinated against B. burgdorferi and/or as having an early, intermediate or chronic B. burgdorferi infection.

10. The method of claim 9, wherein the only antibodies to B. burgdorferi proteins determined in the method are antibodies to B. burgdorferi OspA, OspC, and OspF.

11. The method of claim 10, wherein the mammal is identified as not infected with B. burgdorferi by determining an absence of the antibodies to the OspA, OspC and OspF proteins.

12. The method of claim 10, wherein the determining the presence or the absence of the antibodies is determined using a fluorescent bead-based multiplex assay.

13. The method of claim 10, wherein the reference is a range of values for median fluorescence intensities.

14. A method for diagnosing Lyme disease status in a mammal comprising, in a biological sample obtained or derived from the mammal, determining the presence or absence of antibodies specific for Borrelia burgdorferi (B. burgdorferi) outer surface proteins (Osp) OspA, OspC, and OspF, and:

i) identifying the mammal as vaccinated against but not infected by B. burgdorferi based on determining antibodies to OspA and an absence of antibodies to OspC or OspF;

ii) identifying the mammal as having an early B. burgdorferi infection based on determining antibodies to OspC and an absence of antibodies to OspA and OspF;

iii) identifying the mammal as having chronic B. burgdorferi infection based on determining antibodies to OspF and an absence of antibodies to OspA and OspC;

iv) identifying the mammal as vaccinated against and having an early B. burgdorferi infection based on determining antibodies to OspA and OspC and an absence of antibodies to OspF;

v) identifying the mammal as vaccinated against and having a chronic B. burgdorferi infection based on determining antibodies to OspA and OspF and an absence of antibodies to OspC;

vi) identifying the mammal as having an intermediate B. burgdorferi infection based on determining antibodies to OspC and OspF and an absence of antibodies to OspA;

or

vii) identifying the mammal as having been vaccinated against and having an intermediate B. burgdorferi infection based on determining the presence of antibodies to OspC, OspF and OspA.

15. A composition comprising an isolated protein comprising the sequence of SEQ ID NO:

15, SEQ ID NO;

17, SEQ ID NO;

19, or a combination of said proteins.

16. The composition of claim 15, comprising the combination of proteins comprising SEQ ID NO:

15, SEQ ID NO;

16 and SEQ ID NO;

17.