Patent
Disclosed is the use of isolated Ats-1 protein in Anaplasma phagocytophilum in the ELISA detection of Anaplasma pathogen. The recombinant expression of Ats-1 and its use as a kit for ELISA are also disclosed.
1. A method of detecting the presence of an anti- Anaplasma translocated substrate-1 (Ats-1) antibody against Anaplasma phagocytophilum in a biological sample of a patient suspected of having an infection of Anaplasma phagocytophilum , comprising the steps of:
(i) immobilizing an isolated Ats-1 protein antigen onto a surface of a support;
(ii) contacting said immobilized Ats-1 protein with a biological sample from a patient suspected of containing an anti-Ats-1 antibody against Anaplasma phagocytophilum , under conditions that allow formation of a complex of the antibody and the protein antigen; and
(iii) detecting the formation of said antibody-protein antigen complex,
wherein said detected antibody-protein antigen complex is indicative of the presence of said anti-Ats-1 antibody against Anaplasma phagocytophilum in said biological sample of said patient.
2. The method of claim 1 , wherein said isolated Ats-1 protein is a recombinant protein.
3. The method of claim 1 , wherein said support is selected from the group consisting of polyethylene, polypropylene and glass.
4. The method of claim 1 , wherein said support is a microtiter well.
5. The method of claim 1 , wherein said biological sample is whole blood.
6. The method of claim 1 , wherein said antibody is an IgG.
7. The method of claim 1 , wherein said antibody is an IgM.
8. The method of claim 1 , wherein said method is an ELISA.
9. The method of claim 8 , wherein said ELISA has a sensitivity of at least 90%.
10. The method of claim 8 , wherein said ELISA has a specificity of at least 75%.
11. A method of diagnosing an infection of Anaplasma phagocytophilum in a patient by detecting the presence of an anti-Ats-1 antibody, comprising the steps of:
(i) obtaining a biological sample from a patient suspected of having an anti-Ats-1 antibody against Anaplasma phagocytophilum;
(ii) immobilizing an isolated Ats-1 protein antigen onto a surface of a support;
(iii) contacting said immobilized Ats-1 protein with said biological sample, under conditions that allow formation of a complex of the antibody and the protein antigen;
and
(iv) detecting said antibody-protein antigen complex,
wherein the presence of said detected antibody-protein antigen complex is indicative that said patient has an infection of Anaplasma phagocytophilum.
12. The method of claim 11 , wherein said isolated Ats-1 protein is a recombinant protein.
13. The method of claim 11 , wherein said support is selected from the group consisting of polyethylene, polypropylene and glass.
14. The method of claim 11 , wherein said support is a microtiter well.
15. The method of claim 11 , wherein said biological sample is whole blood.
16. The method of claim 11 , wherein said antibody is IgG.
17. The method of claim 11 , wherein said antibody is an IgM.
18. The method of claim 11 , wherein said method is an ELISA.
19. The method of claim 18 , wherein said ELISA has a sensitivity of at least 90%.
20. The method of claim 18 , wherein said ELISA has a specificity of at least 75%.