Methods, devices, kits and compositions for detecting roundworm, whipworm, and hookworm

• US 8,895,294 B2
• Filed: 03/07/2013
• Issued: 11/25/2014
• Est. Priority Date: 06/15/2007
• Status: Active Grant

• Current Assignee: Monsanto Company (Bayer AG), IDEXX Laboratories Incorporated
• Original Assignee: Divergence, Inc. (Bayer AG), IDEXX Laboratories Incorporated
• Inventors: Elsemore, David Allen, Geng, Jinming, Flynn, Laurie A., Crawford, Michael
• Primary Examiner(s): NAVARRO, ALBERT MARK
• Application Number:
  US13/788,532
• Publication Number:
  US 20140205994A1
• Time in Patent Office: 628 Days
• Field of Search: None
• US Class Current: 435/287.1
• CPC Class Codes: G01N 2333/43526 from worms
• CPC Class Codes: G01N 2333/4353 from nematodes
• CPC Class Codes: G01N 2469/10 Detection of antigens from microorganism in sample from host
• CPC Class Codes: G01N 33/5308 for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
• CPC Class Codes: G01N 33/56966 Animal cells
• CPC Class Codes: G01N 33/6893 related to diseases not provided for elsewhere

Abstract

Methods, devices, kits and compositions for detecting the presence or absence of one or more helminthic coproantigens in a sample are disclosed herein. The methods, devices, kits and compositions of the present invention may be used to confirm the presence or absence of roundworm, whipworm and/or hookworm in a fecal sample from a mammal and may also be able to distinguish between one or more helminth infections. Confirmation of the presence or absence of roundworm, whipworm and/or hookworm in the mammal may be made, for example, for the purpose of selecting an optimal course of treating the mammal and/or for the purpose of determining whether the mammal has been rid of the infection after treatment has been initiated.

Claims

1. A device for detecting the presence or absence of one or more helminthic antigens from a sample, the device comprising a solid support, wherein the solid support has immobilized thereon two antibodies selected from the group consisting of:

(i) a first antibody capable of specifically binding a roundworm coproantigen, but not a whipworm coproantigen and a second antibody capable of specifically binding a whipworm coproantigen, but not a roundworm coproantigen;

(ii) a first antibody capable of specifically binding a roundworm coproantigen, but not a hookworm coproantigen and a second antibody capable of specifically binding a hookworm coproantigen, but not a roundworm coproantigen; and

(iii) a first antibody capable of specifically binding a whipworm coproantigen, but not a hookworm coproantigen and a second antibody capable of specifically binding a hookworm coproantigen, but not a whipworm coproantigen.

2. The device according to claim 1, further comprising one or more reagents to detect one or more of the group consisting of:

one or more non-worm parasites, heartworm, one or more viruses, one or more fungi, and one or more bacteria.

3. The device according to claim 1, wherein said one or more reagents are one or more antibodies or one or more antigens recognized by antibodies specific for one or more non-worm parasites, one or more viruses, one or more fungi or one or more bacteria.

4. The device according to claim 1, wherein the antibody capable of specifically binding a roundworm coproantigen is capable of specifically binding a polypeptide consisting of an amino acid sequence of SEQ ID NO:

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

15, SEQ ID NO;

16, SEQ ID NO;

19, SEQ ID NO;

20, SEQ ID NO;

21, SEQ ID NO;

22, SEQ ID NO;

23, SEQ ID NO;

26, SEQ ID NO;

27, SEQ ID NO;

28, SEQ ID NO;

29, SEQ ID NO;

30, SEQ ID NO;

38, or Copro6728;

or the first antibody was raised against an extract of whole roundworms, or extract of roundworm reproductive organs, or extract of roundworm intestines.

5. The device according to claim 1, wherein the antibody capable of specifically binding a whipworm coproantigen is capable of specifically binding a polypeptide consisting of an amino acid sequence of SEQ ID NO:

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

8, or SEQ ID NO;

9.

6. The device according to claim 1, wherein the antibody capable of specifically binding a hookworm coproantigen is capable of specifically binding a polypeptide consisting of an amino acid sequence of SEQ ID NO:

33 or SEQ ID NO;

34, or CoproASP5.

7. The device according to claim 1, wherein the sample is obtained from a mammal that is a canine or a feline.

8. The device according to claim 1, wherein the roundworm is Toxocara canis, Toxocara cati, Toxocara vitulorum, Toxascaris leonina, Baylisascaris procyonis, Ascaridia galli, Parascaris equorum, Ascaris suum, Ascaris lumbricoides, Anisakis simplex, or Pseudoterranova decipiens.

9. The device according to claim 1, wherein the whipworm is Trichuris vulpis, Trichuris campanula, Trichuris serrata, Trichuris suis, Trichuris trichiura, Trichuris discolor and Trichocephalus trichiuris.

10. The device according to claim 1, wherein the hookworm is Ancylostoma caninum, Ancylostoma braziliense, Ancylostoma duodenal, Ancylostoma ceylanicum, Ancylostoma tubaeforme, Ancylostoma pluridentatum, Necator americanus, and Uncinaria stenocephala.

11. The device according to claim 1, wherein the first and second antibodies do not specifically bind any coproantigen derived from heartworm.

12. The device according to claim 1, wherein the two antibodies are capable of forming antibody-coproantigen complexes in the presence of one or more helminthic antigens and further including at least one secondary antibody that binds to at least one of the complexes.

13. The device according to claim 12, wherein the at least one secondary antibody is labeled.

14. The device according to claim 1, wherein one or more of the first and second antibodies are labeled.

15. The device according to claim 1, wherein the solid support forms part of an enzyme-linked immunosorbent assay device.

16. The device according to claim 15, wherein the enzyme-linked immunosorbent assay device is a lateral flow immunoassay device.

17. The device according to claim 1, wherein the roundworm coproantigen, hookworm coproantigen, and/or whipworm coproantigen is from a fecal sample.

18. A kit for detection of one or more roundworm coproantigens, one or more whipworm coproantigens and/or one or more hookworm coproantigens in a mammalian sample, the kit comprising the device of claim 1, and one or more reagents sufficient for the detection of the one or more antigens.

19. The kit according to claim 18, wherein the one or more reagents are selected from the group consisting of one or more indicator reagents, one or more antibody labeling compounds, one or more antibodies, one or more antigen capture reagents, one or more inhibitors, and one or more wash reagents.

20. A device comprising a solid support having immobilized thereon a first antibody and a second antibody, wherein the first antibody, the second antibody or both are complexed with a coproantigen and a secondary antibody, and wherein the first antibody and second antibody are a pair selected from the group consisting of:

(i) a first antibody capable of specifically binding a roundworm coproantigen, but not a whipworm coproantigen and a second antibody capable of specifically binding a whipworm coproantigen, but not a roundworm coproantigen;

(ii) a first antibody capable of specifically binding a roundworm coproantigen, but not a hookworm coproantigen and a second antibody capable of specifically binding a hookworm coproantigen, but not a roundworm coproantigen; and

(iii) a first antibody capable of specifically binding a whipworm coproantigen, but not a hookworm coproantigen and a second antibody capable of specifically binding a hookworm coproantigen, but not a whipworm coproantigen.

21. The device of claim 20, wherein at least one antibody in the pair is polyclonal.

22. The device of claim 20, wherein the secondary antibody is labeled.