Characterization of granulocytic ehrlichia and methods of use
The present invention relates, in general, to polypeptides having antigenic epitopes from granulocytic ehrlichia (GE) proteins and methods of use thereof.
1. A polypeptide consisting essentially of an amino acid sequence encoded by a nucleotide sequence contained within nucleotides 232 to 760 of SEQ ID NO: 26, wherein the amino acid sequence encoded by the nucleotide sequence contained within nucleotides 232 to 760 of SEQ ID NO: 26 comprises an antigenic epitope of at least 7 amino acids in length.
2. The polypeptide of claim 1 , wherein the polypeptide consists of an amino acid sequence encoded by a nucleotide sequence contained within nucleotides 232 to 760 of SEQ ID NO:26.
3. The polypeptide of claim 1 , wherein the antigenic epitope comprises the amino acid sequence of SEQ ID NO: 69.
4. The polypeptide of claim 1 , wherein the amino acid sequence is at least 10 amino acids in length.
5. The polypeptide of claim 1 , wherein the amino acid sequence is at least 15 amino acids in length.
6. The polypeptide of claim 1 , wherein the amino acid sequence is at least 20 amino acids in length.
7. The polypeptide of claim 1 , wherein the amino acid sequence is at least 25 amino acids in length.
8. A composition comprising the polypeptide of any one of claims 1 - 3 and 7 .
9. A diagnostic kit comprising: a container that contains the polypeptide of any one of claims 1 to 3 .
10. A diagnostic kit comprising:
(i) a first container that contains the polypeptide of claim 7 ; and
(ii) a second container that contains a detection reagent that detects an immunocomplex comprising the polypeptide bound to an antibody of a subject.
11. The diagnostic kit of claim 10 , wherein the first container further comprises a wash reagent.
12. The diagnostic kit of claim 10 , wherein the subject is a dog, horse, cow, pig, sheep, chicken, or human.
13. The diagnostic kit of claim 10 , wherein the subject is a dog.
14. The diagnostic kit of claim 10 , wherein the detection reagent comprises a radiolabel, an enzymatic label, or a fluorescent label.
15. A method of detecting an antibody to granulocytic Ehrlichia E8 protein in a sample obtained from a subject, comprising:
(a) contacting the sample with the polypeptide of any one of claims 1 to 3 , under conditions in which immunocomplexes form, wherein the immunocomplexes comprise the polypeptide bound to the antibody; and
(b) detecting the presence of the immunocomplexes, thereby detecting the antibody to granulocytic Ehrlichia E8 protein in the sample.
16. A method of detecting an antibody to granulocytic Ehrlichia E8 protein in a sample obtained from a subject, comprising:
(a) contacting the sample with the polypeptide of claim 7 , under conditions in which immunocomplexes form, wherein the immunocomplexes comprise the polypeptide bound to the antibody; and
(b) detecting the presence of the immunocomplexes, thereby detecting the antibody to granulocytic Ehrlichia E8 protein in the sample.
17. The method of claim 16 , wherein, in step (b), detecting the presence of the immunocomplexes comprises mixing the immunocomplexes with a detectably labeled antibody that binds specifically to the antibody in the immunocomplexes.
18. The method of claim 17 , wherein the detectably labeled antibody comprises a radiolabel, an enzymatic label, or a fluorescent label.
19. The method of claim 16 , wherein the sample is tissue, blood, serum, plasma, or urine.
20. The method of claim 16 , wherein the subject is a dog, horse, cow, pig, sheep, chicken, or human.
21. The method of claim 16 , wherein the subject is a dog.